459600

antibody from Invitrogen Antibodies

Targeting: MT-CO1 COI, COX1, MTCO1

Provider product page for 459600

Western blot

Immunocytochemistry

Immunohistochemistry

Flow cytometry

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Antibody data

Antibody Data
Antigen structure
References [110]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [1]
Other assay [43]

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Product number: 459600 - Provider product page
Provider: Invitrogen Antibodies
Product name: MTCO1 Monoclonal Antibody (1D6E1A8)
Antibody type: Monoclonal
Antigen: Other
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: 1D6E1A8
Vial size: 100 µg
Concentration: 1.0 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

MT-CO1 protein structure - 459600 shown in red.

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Experimental details: Figure 3 GRSF1 Is Required for Mitochondrial Gene Expression (A) Left panel, immunoblot analysis of 143B cells treated with control (Ctrl) or GRSF1 RNAi1. Right panel, immunoblot analysis of 143B cells infected with lentiviruses carrying an empty vector (pLKO) or a shRNA (RNAi2) against GRSF1. COX1 and COX2 are mitochondrially encoded proteins. SDHB and Tom20 are nuclear-encoded mitochondrial proteins. Actin is a nuclear-encoded cytosolic protein. (B) Acidification of the culture medium in GRSF1 RNAi1-treated 143B cells. The number of cells was the same in both conditions. Similar results were obtained in GRSF1 RNAi2-infected cells (data not shown). (C) NanoString analysis of 11 mitochondrially encoded ORFs in control (Ctrl) or GRSF1 RNAi1-treated 143B cells. RNA14 corresponds to bicistronic MTATP8 - MTATP6 RNA. Data are shown as mean +- SEM (n = 3). (D) NanoString analysis of GRSF1 in control (Ctrl) or GRSF1 RNAi1-treated 143B cells. Data are shown as mean +- SEM (n = 3). (E) NanoString analysis of 11 nuclear-encoded proteins in control (Ctrl) or GRSF1 RNAi1-treated 143B cells. Data are shown as mean +- SEM (n = 3). (F) Northern blot analysis of 12S and 16S rRNA in control (Ctrl) or GRSF1 RNAi1-treated cells. TUB, tubulin. (G) Phosphorimager quantification of (E). Data are shown as mean +- SEM (n = 3). (H) 35 S-labeling of mitochondrial translation of 143B cells infected with lentiviruses carrying an empty vector (pLKO) or a shRNA (RNAi2) against GRSF1. (I) Phosphorimager qu

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Experimental details: Fig. 1 Exosomes shuttle mitochondrial proteins. a Differential ultracentrifugation protocol for isolating and purifying exosomes from cell culture supernatants. b Size distribution analysis by Nanoparticle Tracking Analysis (NTA) of purified EVs from primary human T lymphoblasts. c Gene ontology (GO) cellular component analysis of peptides identified in T cell EVs. d Western blot analysis of mitochondrial proteins in EVs obtained from the culture supernatant of primary human T lymphoblasts with or without treatment with phorbol myristate acetate (PMA) plus ionomycin. Cells and EVs were blotted for proteins associated with mtDNA (TFAM and ATAD3), for proteins located in the inner mitochondrial membrane (COX1 and Cytochrome C), the outer mitochondrial membrane (VDAC1), the mitochondrial matrix (mitochondrial manganese superoxide dismutase, MnSOD), and for the exosome markers TSG101 and CD81. e Western blot analysis of ATAD3, TFAM, and the exosome markers CD81 and TSG101 in sucrose fractions. EVs obtained from the culture supernatant of human T lymphoblasts were laid on a discontinuous sucrose gradient and floated by overnight centrifugation. Gradient fractions were collected and analyzed by immunoblot to reveal the distribution of mtDNA-binding proteins and exosomal proteins in the sucrose fractions from lower to higher sucrose density (left to right). Gels shown are representative out of three independent experiments

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Experimental details: Fig 1 Characterization of TFAM and Twinkle (TW) mice. (A) Expression of human TFAM (hTFAM) in left ventricle (LV) and aorta in TFAM and wild-type (WT) control mice. (B) mtDNA copy number in myocardium from TFAM and TW mice by real-time PCR (n = 4). (C) Expression of endogenous murine TFAM (mTFAM) and mitochondrial complex proteins in LV of TFAM and TW mice. (D) Transcription of mtDNA-encoded genes in TFAM and TW mice (n = 6). Data are expressed as mean +- SEM. * P < 0.05 vs. WT, ** P < 0.01 vs. WT, analyzed by Student's t- test.

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Experimental details: FIGURE 8. FAHD1 immunofluorescence. Confocal images of immunofluorescence staining of HUVEC for FAHD1 ( green , left column ), three different localization markers ( red , center column ), and the resulting merge ( right column ) are shown. Complex II and complex IV were used as mitochondrial markers, and catalase was used as peroxisomal marker.

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Experimental details: Figure 5 Representative blots (upper panel) and quantitative analysis of the expression (lower panel) of arginase I in the cytosolic fraction (A) and arginase II in the mitochondrial fraction (B) of myocardial samples from the non-ischemic and ischemic area of pigs subjected to ischemia-reperfusion and myocardium from sham operated pigs. Expression of arginase I is normalized to that of the surface membrane L-type Ca 2+ channel, dihydropyridine receptor (DHPR) and arginase II to that of cytochrome oxidase (COX) and presented in % of the mean expression in the sham group. Mean+-SEM, n = 6. There were no significant differences between the groups.

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Experimental details: Figure 1 Decreased spontaneous cell death and caspase signaling but constitutive autophagy in memory B cells ( a ) Percentages of cell loss of NP- or HA-specific memory or GC B cells after in vitro culture. Experiments were performed four times in triplicates using cells from a pool of 15 mice. ** P

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Experimental details: Figure 4 Loss of memory B cells in the absence of Atg7 ( a , b ) DUMP - B220 + IgG1 + CD38 + NP + memory B cells in the spleen of B/ Atg7 -/- mice or WT controls eight weeks after immunization. Unimmunized mice were used as negative controls (a). Total NP-specific memory B cells in the spleen are presented in (b). ** P

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Experimental details: Figure 6 Nuclear CLK-1/COQ7 suppresses the expression of a subset of UPR mt genes (a) CLK-1 nuc(+) expressing worms display decreased hsp-6::gfp reporter activity compared to clk-1(-) worms. The unmodified hsp-6::gfp reporter strain is designated clk-1(+) . Quantification of reporter fluorescence in CLK-1 nuc(+) expressing worms ( clk-1(-); clk-1 nuc(+) ) relative to clk-1(-) worms (mean fluorescence of 50 worms per genotype pooled from n=3 independent experiments; error bars, s.e.m. ** P < 0.005). (b) qPCR measuring mRNA transcripts of UPR mt genes in clk-1(-) or clk-1(-); clk-1 nuc(+) worms relative to wild type strain ( N2 ) (mean values from 3 reactions per condition in n=3 independent experiments; error bars, s.e.m., n.s., no significant difference, * P < 0.05). (c) Heat map depicting change in expression of UPR mt genes (MT) and UPR ER (ER) genes in R28A cells compared to WT COQ7 cells. Map generated from the qPCR data presented in Supplementary Figure 4b and is representative of n=3 independent experiments. Scale represents mean fold change in expression. (d) Immunoblots of levels of UPR mt proteins including the mitochondrial controls COXIV (nuclear-encoded) and MTCO1 (mitochondrial-encoded). Uncropped images of blots are shown in Supplementary Fig. 5 .

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Experimental details: Fig 2 Rotenone induces delayed Parkin translocation to mitochondria in PINK1-/- myocytes. (A) The mitochondrial complex I inhibitor rotenone (40 muM) significantly reduced mitochondrial membrane potential by 1 hour of treatment in WT and PINK1-/- cardiac myocytes (n = 3). (B) Quantitation of percentage of cells with Parkin translocation to mitochondria after 60 and 90 min of rotenone treatment. (C) Representative images. Mean +- SEM, n = 3, *p

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Experimental details: Fig 1 Translocation of Parkin to mitochondria and activation of mitophagy occur independently of PINK1. (A) Perfusion of WT and PINK1-/- hearts with the mitochondrial uncoupler FCCP (100 nM) led to accumulation of Parkin in the mitochondrial fraction within 15 minutes (15'). (B) Perfusion of WT hearts with FCCP for up to 15 minutes did not result in accumulation of PINK1 in the mitochondrial fraction. PINK1-/- perfused heart samples shown for comparison. In vivo FCCP treatment for 24 h led to (C) translocation of Parkin to cardiac mitochondria, (D) increased ubiquitination of cardiac mitochondrial proteins, and (E) increased LC3II association with mitochondria. (F) Analysis of mitochondria isolated from the infarct border zone four hours after in vivo myocardial infarction (MI). Parkin levels increased in the mitochondrial fraction in both WT and PINK1-/- hearts after MI. Representative western blots and densitometry data show significantly elevated Parkin protein levels in the heart (G) and at the mitochondria (H) in 3 month old PINK1-/- mice. Mean +- SEM, n = 3, *p

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Experimental details: Figure 3 Representative images showing SNAP23 colocalization with the plasma membrane, mitochondria, and IMTG. Representative images showing SNAP23 colocalization with the plasma membrane marker dystrophin (A), mitochondrial marker COX (B), and IMTG staining using oil red O (C). All images were obtained using a 63x oil objective of a confocal microscope. Bar = 30 mu m. Pearson's correlation coefficients are shown on the merged images (mean +- SEM). The Pearson's correlation coefficients of the nonmatched pairs of images were; SNAP23 and Dystrophin ( r = 0.007 +- 0.003); SNAP23 and IMTG ( r = 0.000 +- 0.003); SNAP23 and COX ( r = 0.006 +- 0.005). All these values were significantly lower ( P < 0.05) than the matched image pairs demonstrating that the colocalization detected was not due to chance. The bottom panels of images highlight specific regions of interest which best visualize the overlapping yellow pixel clusters (i.e., colocalization) in plasma membrane, LDs, and mitochondria.

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Experimental details: Fig. 2 Ab93374 (Abcam) recognizes MPV17 specific structures in U2OS cells transiently transfected with an MPV17 expression construct in immunofluorescence analysis. Ab93374 signal ( green ) is largely missing in nontransfected cells. The immunoreactivity does colocalise with peroxisomal (anti-catalase, mouse polyclonal, Abcam ab88650) and mitochondrial (anti-complex IV mitochondrial subunit I, mouse monoclonal, Invitrogen 459600) markers ( red ). Immunofluorescence (IF) method was according to Pircher et al. [ 16 ] using a MicroRadiance confocal scanning system (Bio-Rad) in combination with a Zeiss Axiophot microscope. Colocalisation analysis was performed using the Fiji software of Image J and is illustrated by white dots

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Experimental details: Fig 6 Loss of the m -AAA protease in oligodendrocytes causes mitochondrial dysfunction and dark cell death. (A) Deconvoluted single-plane confocal images of double immunofluorescence staining of mt-YFP and COX1 in WT and DKO mice at the indicated age. mt-YFP + mitochondria in DKO lose COX1 staining. Scale bar, 1 mum. (B) Deconvoluted single-plane confocal images of double immunofluorescence staining of mt-YFP and cytochrome c in WT and DKO mice at 8 weeks. Scale bar, 1 mum. (C) Ultrastructural analysis shows an oligodendrocyte undergoing dark cell death in the corpus callosum of 8-week-old DKO mice. M: mitochondria; N: nucleus. Scale bar, 1 mum. n = 3 per genotype in all experiments.

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Experimental details: Figure 4 Expression of arginase II in the cytosolic and mitochondrial fractions of a representative myocardial sample. For comparison the expression of the mitochondrial membrane protein cytochrome oxidase (COX) is depicted.

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Experimental details: Figure 3 Representative Western blots of Complex I-V, Cytochrome C, Pyruvate Dehydrogenase (PDH) subunit E1alpha protein, PDH site1 and PDH site2 phosphorylation and ACC Ser211 phosphorylation.

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Experimental details: Figure 3 Oxidative stress-dependent translocation of DJ-1 into mitochondria. Representative confocal micrographs of mouse primary RPE (A-F) and ARPE-19 (G-L) monolayers plated on Transwells(r) and labeled with antibodies to DJ-1 (A, D, G, J) and COX IV (B, E, H, K). Under baseline conditions, there is very little colocalization between DJ-1 and COX IV, as observed in overlaid images (C, I) for both RPE cultures; DJ-1 is mostly distributed trough the cytoplasm (arrows) and to the nuclei (*) of some cells. Upon oxidative stress induced by incubation with 400 uM H 2 O 2 for 1 hr, DJ-1 staining is increased both in the mouse primary (D) and ARPE-19 (J) cultures. In cultures treated with H 2 O 2 some DJ-1 re-distributed to mitochondria (arrowheads) and displayed significant colocalization with COX IV in overlaid images (F, L). Scale bar = 10 um.

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Experimental details: Figure 1 Exercise training increase mitochondrial oxidative phosphorylation complexes in an AMPK alpha2-dependent manner . Protein levels of oxidative phosphorylation complexes, (A) Complex I, (B) Complex II, (C) Complex III, (D) Complex IV, (E) Complex V (oligomycin-sensitivity conferring protein subunit, OSCP), and (F) Cytochrome C were evaluated in quadriceps muscle of sedentary (control) or exercise trained female WT and AMPK alpha2 KD mice ( n = 13-15). Values are mean +- SEM. * indicates vs. WT control ( p < 0.05) analyzed by unpaired t -tests.

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Experimental details: Figure 4 Repeated treatment with AICAR increases abundance of mitochondrial electron transport chain proteins in an AMPK alpha2-dependent manner . WT and AMPK alpha2 KD male mice were given daily subcutaneous injections with AICAR (500 mg/kg body weight) or saline for 4 weeks. (A-E) Protein abundance of oxidative phosphorylation Complexes I-V and (F) Cytochrome C was measured in quadriceps muscle ( n = 7-8). A significant interaction effect (treatment x genotype; p < 0.05) was observed for Cytochrome C. Values are mean +- SEM. * indicates vs. WT saline ( p < 0.05) analyzed by t -tests, ** indicates vs. WT saline ( p < 0.01), and ++ indicates genotype effect within AICAR treated animals ( p < 0.01).

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Experimental details: Figure 1 MQC markers increase and mitochondrial content decreases in spinal cord of symptomatic SOD 1-G93A mice A, B Representative Western blot (A) and quantification (B) of p62 associated with mitochondria from spinal cords. Protein levels are normalized by subunit ATPase beta of mitochondria (Complex V). Mitochondrial p62 levels are increased at 120 days (symptomatic stage) in SOD1-G93A spinal cord relative to Non Tg and wild-type SOD1 (wtSOD1). Results are expressed as mean +- SEM relative to Non Tg at 30 days; n = 8 mice (four males and four females). At 120 days, ** P = 0.0018 (Non Tg vs. SOD1-G93A) and ** P = 0.0011 (SOD1-G93A vs. wtSOD1) by paired one-way ANOVA with Tukey's correction. No other statistically significant differences were found (paired Friedman's test with Dunn's correction at 30 days and paired one-way ANOVA with Tukey's correction at 60 and 90 days). C, D Representative Western blot (C) and quantification (D) of Tim23 in homogenates from spinal cords using beta-actin as normalizer. Results are expressed as mean +- SEM and relative to Non Tg at 30 days; n = 6 mice (three males and three females). *** P = 0.0002 (Non Tg vs. SOD1-G93A at 120 days) by paired one-way ANOVA with Tukey's correction. No other statistically significant differences were found (paired Friedman's test with Dunn's correction at 30 days and paired one-way ANOVA with Tukey's correction at 60 and 90 days). Tim23 is decreased in SOD1-G93A spinal cord relative to Non Tg

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Experimental details: Figure 1 Western blot analyses of distinct mitochondrial proteins in lung homogenates from newborn, P15, and adult mice. (a) Following the homogenization, 50 mu g of protein isolated from the lungs of each group (NB, P15, and adult) was resolved by 10% SDS-polyacrylamide gel electrophoresis, and the blots were immunostained with anti-MT-ND1, anti-SDHD, anti-COX1 (OXPHOS complex IV subunit I), anti-COX2 (OXPHOS complex IV subunit II), anti-ATP synthase (ATP5b), and anti-POLG2. GAPDH was used as a loading control. * Nuclear-encoded protein. ** Mitochondrially encoded protein. (b) Bar graphs summarizing normalized data. p values were calculated by the one-way ANOVA using Tukey's test. n = 3; * p

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Experimental details: Fig 3 Defective mitochondrial biogenesis upon CTCF loss. Western blot of Cox I, Cox IV (A), Grp75, Tfam and Tom20 (B) of control and mutant ( Ctcf fl / fl ; Nkx2-5-Cre ) hearts at E10.5 and E11.5. (C) Blue native gel of control and mutant hearts at E10.5 and E11.5 showing abundance and distribution of mitochondrial respiratory complexes (CI, CIV, CVm) and supercomplexes (CI+CIII). (D) Densitometry of blue native gel quantifying changes between E10.5 and E11.5 in complexes and supercomplexes, in control and mutant hearts. (E) Transmission Electron Microscopy at E10.5 and E11.5 in control and mutant cardiomyocytes. n, nucleus; sm, sarcomere. Arrowheads point to mitochondria. Scale bar, 1 mum top row, 0.5 mum bottom row.

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Experimental details: Fig. 5 Increase of mitochondrial size and dysfunction, oxidative stress and impaired SIRT1 expression and autophagic activities. a Representative transmission electron (TEM) micrographs showing the mitochondria located in the IHCs and OHCs from WT and p43 -/- mice aged 6 months. Note the increase in the number of mitochondria with the lack of their cristae (arrowheads) and enlarged mitochondria in the hair cells of p43 -/- mice. Scale bars = 0.5 mum. b The histogram represents the mean diameter of the mitochondria in the IHCs and OHCs from p43 -/- (red bars) and WT (blue bars) mice obtained by TEM imaging measurements. Data are expressed as mean +- SD (n = ~ 45 to 50 mitochondria, taken randomly from the IHCs or OHCs, 4 cochleae per age and strain). One-way ANOVA was followed by Dunn's test (* P = 0.014, *** P

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Experimental details: Immunocytochemical visualization of stained reaction with antibodies against ( a ) MTCO-1 (Cytochrome c oxidase subunit I) and ( b ) 8-OHdG (8-hydroxy-2'-deoxyguanosine) in human gastric cancer cells sensitive (EPF85-237 P) and chemoresistant (EPF85-237 RDB) after the 72 h exposure to normal (NG) and altered gravity (MG) with doxorubicin (DOX = 0.5 ng/mL). Scale bars correspond to 50um.

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Experimental details: Overexpression of mt-ELP3 results in an increase in the expression of mitochondrial OXPHOS complexes and mitochondrial respiration. ( A ) and ( C ) Representative BN-PAGE gel stained with Coomassie Brilliant Blue of respiratory complexes purified from control cells (pcDNA T0) and cells overexpressing mt-ELP3 (pcDNA mt-ELP3) ( A ), and from control cells (pcDNA T0) and cells overexpressing mt-ELP3 mutant (pcDNA mt-ELP3 mut) ( C ). A 20-ug aliquot of each sample was loaded per well. A 5-uL aliquot of NativeMark unstained protein standard (Marker) was loaded. ( B ) and ( D ) Representative Blue Native-PAGE of OXPHOS complexes in HEK293T cells transfected with pcDNA4 T0 or pcDNA mt-ELP3 ( B ) and with pcDNA4 T0 or pcDNA mt-ELP3 mut ( D ). The scatter plots show the densitometric measurements of OXPHOS complexes normalized to complex-II (loading control) and represented as fold change relative to control cells. ( E ) Representative immunoblots of mitochondrial OXPHOS subunits ND1 (Complex I), MTCO1 (Complex IV), CytB (Complex III), ATP5A (Complex V) and SDHA (Complex II) in HEK293T cells transfected with pcDNA4 T0, pcDNA mt-ELP3 or pcDNA- mt-ELP3 mut. The membranes were also probed with an antibody against ELP3 and b-actin, B-actin was used as a loading control. ( F ) and ( G ) Oxygen consumption rate (OCR). Equal numbers of HEK293T cells transiently transfected with pcDNA T0 or pcDNA mt-ELP3 were subjected to oxygen consumption measurements in a Seahorse XF96 extracellular flux a

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Experimental details: Figure EV4 Increased mitochondrial protein turnover in G93A mice is attenuated by Parkin knockout A, B Western blots of COXI (A) and Tim23 (B) in spinal cord homogenates at 130 days. C Quantification of COXI at 130 days with beta-actin as a normalizer. Results are expressed as mean +- SEM and as percent of Non Tg; n = 8 (four males and four females) mice per group. No statistically significant differences were found between G93A and PKO/G93A ( P = 0.493 by paired Student's t -test). D Quantification of Tim23 at 130 days, using beta-actin as loading control, showed decreased levels of Tim23 in G93A mice. Results are expressed as mean +- SEM and as a percent of Non Tg; n = 8 (four males and four females) mice per group. No statistically significant differences were found between G93A and PKO/G93A ( P = 0.921 by paired Student's t -test). * P = 0.035 (Non Tg Vs. G93A) by paired Friedman's test with Dunn's correction. No other statistically significant differences were found. E, F Representative Western blots of COXI (E) and Tim23 (F) in disease end-stage homogenates. G Quantification of COXI at disease end stage using beta-actin as loading control. Results are expressed as mean +- SEM and as percent of Non Tg; n = 5 (three males and two females) mice per group. * P = 0.047 for (G93A and PKO/G93A) by paired Student's t -test; ** P = 0.0018 by paired Friedman's test with Dunn's correction (Non Tg vs. G93A). H Quantification of Tim23 with beta-actin as normalizer

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Experimental details: Figure 1 CHCHD 10 resides in the same complex as mitofilin, CHCHD 3 and CHCHD 6 Second dimension of the blue native ( BN )- PAGE showing that CHCHD 10 migrates with the MICOS protein subunits mitofilin, CHCHD 3 and CHCHD 6 in isolated mitochondria from mouse brain. BN - PAGE of the MICOS and OXPHOS complexes in control (C) and patient (P1, P2) fibroblasts. Complexes I to IV of OXPHOS were detected with the 39 kDa subunit antibody ( CI ), 70 kDa subunit antibody ( CII ), core I antibody ( CIII ), and cytochrome c oxidase subunit I antibody ( CIV ). MICOS complex was detected with an anti-mitofilin antibody. Second dimension of the BN - PAGE showing that the steady-state levels of assembled CHCHD 10 in MICOS complex are decreased in patient fibroblasts. The dividing lines correspond to gel sections visible in the raw data file. Analysis of OXPHOS supercomplexes in control and patient fibroblasts. BN - PAGE from isolated mitochondria permeabilized with 6 g/g (w/v) of digitonin immunoblotted on PVDF membrane and incubated with the indicated antibodies. SC , supercomplexes I+ III 2 + IV n . The dividing lines correspond to gels sections visible in the raw data file. Source data are available online for this figure.

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Experimental details: Fig. 2 Colonic mitochondriopathy with a robust gene signature for reduced rectal mitochondrial energy functions in UC. a Thirteen mitochondrial-encoded genes are downregulated in UC vs. control with their fold change, FDR corrected P value, and associated mitochondrial complex as indicated. High-Resolution Respirometry was performed on fresh colon biopsies (5 control, 9 with active UC, and 9 with inactive UC) using the Oroboros O2k modular system to evaluate the activity of Complex I ( b ) and Complex II ( c ) of the electron transport chain. JC1 staining and FACS analysis were used to define the mitochondrial membrane potential of d EpCAM + epithelial cells and e CD45 + leukocytes isolated from colon biopsies (7 controls, 6 active UC, and 7 with inactive UC, 85-99% viability). Colon PPARGC1A (PGC-1alpha) expression for f PROTECT cohort, h RISK cohort in (transcripts per million (TPM) values), and for j adult UC cohort (GSE59071 12 ) in normalized values was plotted after stratifying the samples as indicated. g , i , k Krebs cycle TCA gene signature PCA PC1 for the above cohorts is plotted, samples are stratified as indicated. l Representative rectal MT-CO1 and COX5A immunohistochemistry (complex IV) for Ctl ( n = 14), inactive ( n = 10), and active UC ( n = 11) with moderate Mayo endoscopic subscore and moderate PUCAI. Scale bar represents 50 mum. m Frequency of MT-CO1-positive and COX5A-positive epithelial cells out of the total epithelial cells for

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Experimental details: Fig. 4 Mitochondrial alterations in EBS-MD muscle. a Skeletal muscle specimens from a healthy control and EBS-MD patients were histologically double-stained for SDH and COX. Note the presence of ""rubbed-out"" areas ( arrows ), and the presence of COX-negative fibers in patient 3 ( arrowheads ). Scale bar: 50 mum. b Confocal imaging of mitochondrial respiratory complex IV-stained skeletal muscle specimens from a healthy control and EBS-MD patients. Panels i-iv are magnifications of the boxed areas in panel b. Note the reduced staining intensity in all EBS-MD samples. Scale bars: 50 mum ( b ), 25 mum (panels i-iv). c Immunoblotting of cell lysates prepared from EBS-MD patients and three healthy controls using antibodies for mitochondrial respiratory complex II or V. alpha-Actinin was used as loading control. d Signal intensities of respiratory complex II or V protein bands as shown in ( c ) were densitometrically measured and normalized to the total protein content (assessed by alpha-actinin staining). Healthy controls ( dashed line ) are set to 100 %. Mean values +- SEM, 3 replicates

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Experimental details: Figure 1 Cultured and Labelled mesenchymal stem cells morphology by light microscope without superparamagnetic iron oxide (SPIO) labeling (x40) ( A ) and with SPIO labeling after staining with Prussian blue (x40) ( B ). Transmission electron microscopic examination ( C ) x5600 & ( D ) x11000, showing labeled cells with iron particles appear as black aggregates among numerous round endosomes within the cytoplasm (arrows). The nucleus (N) is convoluted, with prominent euchromatin and peripheral heterochromatin, a conspicuous nucleolus (Nu) is also seen. Homing of the SPIO-labelled transplanted MSCs into their niche evident by immunohistochemical (IHC) staining ( E ) & Prussian blue staining ( F ) of the recipient liver where IHC using the anti-OxPhos complex IV subunit I monoclonal antibody showing areas that were negative for cytochrome C oxidase located either in close proximity to, or in direct contact with, the portal tract region (arrows; X40 magnification). Prussian blue staining showing bluish iron particles distributed intracellularly in the same region as the cytochrome C oxidase-deficient patches (arrows; X40 magnification). Successful macrophage depletion from the liver shown by IHC, the normal distribution of Kupffer cells is evident in nondepleted liver ( G ) with significant reduction after using clodronate liposome with very few positive anti f4/80 ab cells in the hepatic parynchyma x40 ( H ). ( I ) Comparison between all fibrosis and all cirrhotic groups regardi