PA1-823A

antibody from Invitrogen Antibodies

Targeting: PPARD FAAR, NR1C2, NUC1, NUCII

Provider product page for PA1-823A

Western blot

Immunocytochemistry

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [24]
Comments [0]
Validations
Immunocytochemistry [1]
Flow cytometry [1]
Other assay [15]

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Product number: PA1-823A - Provider product page
Provider: Invitrogen Antibodies
Product name: PPAR delta Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: PA1-823A detects peroxisome proliferator activated receptor (PPAR) delta from rat, human and mouse cells. This antibody does not detect PPAR alpha or PPAR gamma. PA1-823A has been successfully used in Western blot and immunohistochemical and immunofluoresence procedures. By Western blot, this antibody detects an ~49 kDa protein representing PPAR delta from NIH-3T3 cell lysate. Immunolocalization yeilds predominant nuclear staining. The PA1-823A immunogen is a synthetic peptide corresponding to residues M(1) E Q P Q E E T P E A R E E(14) C of mouse PPAR delta. This sequence is 86% conserved in human PPAR delta. PA1-823A immunizing peptide (Cat. # PEP-026) is available for use in neutralization and control experiments.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

PPARD protein structure - PA1-823A shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Extended Data Figure 6 PPAR-d is the predominant PPAR family member expressed in intestinal progenitors and mediates the effects of HFD a , PPAR-d is the most abundant PPAR family member in ISCs (Lgr5-GFP hi ) and progenitors (Lgr5-GFP low ) based on RNA-seq data. b , Confirmation of PPAR family member mRNA expression levels in ISCs (Lgr5-GFP hi ) and progenitors (Lgr5-GFP low ) by qRT-PCR ( n =5). c , Genes upregulated in HFD ISCs (Lgr5-GFP hi ) versus control ISCs were enriched in PPAR and LXR/RXR motifs. d , GSEA of RNA-seq data identified enrichment of PPAR-d targets in ISCs (Lgr5-GFP hi ) and progenitors (Lgr5-GFP low ) with a HFD. e , Confirmation of induced PPAR-d target gene expression in flow sorted ISCs (Lgr5-GFP hi ) and progenitors (Lgr5-GFP low ) by qRT-PCR ( n= 5). All fold changes were significant with P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Extended Data Figure 7 HFD and PPAR-d signaling boost nuclear beta-catenin localization and activity in intestinal progenitors a , b , HFD-derived ISCs ( a , Lgr5-GFP hi ) and progenitors ( b , Lgr5-GFP low ) required less Wnt3a and R-spondin to initiate organoids compared to C ISCs as measured by comparing organoid-formation in complete ENRW media, which includes EGF, Noggin, R-spondin, and Wnt3a, versus EN media, which includes EGF and Noggin but lacks Wnt3a and R-spondin ( n =3). C-derived progenitors, in contrast to HFD-derived progenitors, rarely formed organoids in either ENRW or EN media. c-f , HFD increased nuclear beta-catenin localization in flow sorted ISCs and progenitors from HFD ( c , n =5) and GW-treated ( d , n =4) mice as determined by immunofluorescence (red=DAPI, cyan=non-phosphorylated beta-catenin, CST 8814S). At least 100 cells per sample were quantified. Representative images are shown in ( e , f ). g - i , HFD ( g ) and GW ( h ) treatment increased the numbers of ISCs and progenitors with beta-catenin + nuclei as assessed by immunostains ( n =4 each). Representative images are shown in ( i ); arrowheads indicate representative nuclear beta-catenin in ISCs (red) and progenitors (black). j - l , Association of PPAR-d and beta-catenin in C and HFD derived intestinal crypts as shown by immunoprecipitation (IP) ( n=3 ). For western blot source data, see Supplementary Figure 1 . (Unless otherwise indicated, data reflect mean +- s.d. from n independent experi

Supportive validation

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Experimental details: Fig. 8 ACOX2 inhibits tumor through the PPARalpha signaling pathway. A The GSEA showed that PPAR ( P = 0.004) and FATTY_ACID_METABOLISM ( P = 0.006) were significantly changed. B The PPI network of ACOX2, PPARA, PPARD, and PPARG was constructed by the String database. C The expression of PPARA, PPARD, and PPARG genes in liver cancer tissues and adjacent noncancerou tissues by using RT-qPCR. D The mRNA expression of PPARA, PPARD, and PPARG was detected by RT-qPCR in HepG2 and SMCC-7721 (NC, the normal HCC cell lines; ACOX2-Ov, overexpression of ACOX2; Sh- ACOX2, knockdown ACOX2). E The protein levels of PPARA, PPARD, and PPARG were detected by Western blot in HepG2 and SMCC-7721 (NC, the normal HCC cell lines; ACOX2-Ov, overexpression of ACOX2; Sh- ACOX2, knockdown ACOX2, * P < 0.05; ** P < 0.01; n.s. not significant). F The Pearson correlation analysis between ACOX2 and PPARA, PPARD, and PPARG by the GEPIA database.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 9 Tempol effects on the activators of mitochondrial biogenesis and oxidative phosphorylation. (A) Western blotting analysis of peroxisome proliferator-activated receptor delta (PPARdelta) and peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha) in the diaphragm (DIA) muscle from the sedentary mdx mice ( mdx Sed), exercise trained mdx mice ( mdx Ex), and exercise trained mdx mice treated with Tempol ( mdx Ex+T). (B) The graphs show the PPARdelta and PGC-1alpha levels in the crude extracts of DIA from the mdx Sed, mdx Ex, and mdx Ex+T groups. (C) Western blotting analysis of the OXPHOS complexes (complex V, III, IV, II, and I) in the DIA muscle from the mdx Sed, mdx Ex, and mdx Ex+T groups. (D) The graphs show the OXPHOS protein in the crude extracts of DIA from the mdx Sed, mdx Ex, and mdx Ex+T groups. The blots of the proteins (top row) and beta-actin (loading control, bottom row) are shown. The intensities of each band were quantified and normalized to those of the corresponding control. All values expressed as mean +- standard deviation (SD). * P

Supportive validation

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Experimental details: Fig. 6 LEF1 regulates lipid metabolism in MYC-expressing cells. a Heatmap of acylcarnitines affected by LEF1 siRNA in myc-/- + MYC fibroblast, generated by metabolomics analyses of cells transfected with either control or LEF1 siRNA. b Rat1 myc-/- fibroblasts expressing either vector or human MYC were collected and subjected for RNA-seq analysis and expression levels of genes involving lipid metabolism were compared. c Rat1 myc -/- control fibroblast cells and Rat1 myc -/- cells reconstituted with human MYC were transfected with control or LEF1 siRNAs for 3 days and extracted with NP40 lysing buffer for Western blot assays with the indicated antibodies. d HCEC expressing truncated APC were transfected with either control or LEF1 siRNA for 3 days, and then fractionated into nuclear and cytoplasmic fractions for Western blotting with the indicated antibodies. e DLD1 cells transfected with control of LEF1 siRNAs for three days were extracted with NP40 lysis buffer, and the total lysates were subjected to Western blot assays. f ACAD9 and CADS gene expression in colon cancer and adjacent normal tissues deposited in the TCGA database. Blue: downregulated (< 0.7), grey: no change (0.7-1.3), and red: upregulated (> 1.3). n = 41 pairs. g Human colon tumor tissues (T) and their surrounding benign tissues (B) were extracted with NP40 lysis buffer, and the total lysates were subjected to Western blotting with the indicated antibodies. h Model of MYC and LEF1 positive feedback loop in pro

Supportive validation

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Experimental details: Figure 7 Direct effects of beta-RA on adipogenesis. ( A ) Percentage of 3TL1 cells after seven days of treatment with 1 mM beta-RA relative to the number of untreated 3TL1 cells. Cells cultured in proliferative conditions. ( B ) Levels of the proteins SKP2, p27, and CYCA2, which were involved in the control of the cell cycle. The 3TL1 cells were treated for seven days with 1 mM beta-RA in proliferative conditions. ( C ) Levels of the proteins SKP2, p27, and CYCA2, which were involved in cell cycle control. The 3TL1 cells were treated for seven days with 1 mM beta-RA in differentiative conditions. ( D , E ) Oil Red O staining in 3TL1 cells cultured under proliferative ( D ) and proliferative + differentiative ( F ) conditions. The 3TL1 cells were treated with 1 mM beta-RA from day 0 in both conditions and the stains were performed on three different days (2, 4, and 7). ( F ) Percentage of the area corresponding to the Oil Red O stains in the 3TL1 cells in differentiative conditions after days 4 and 7 of treatment with 1 mM beta-RA. ( G , H ) Levels of PPARgamma and PPARdelta in the 3TL1 cells cultured in proliferative + differentiative ( F ) conditions and treated with 1 mM beta-RA. The results in non-differentiated cells are shown in line one as the negative control. ( I , J ) Levels of CoQ 9 in the 3TL1 cells cultured in proliferative conditions ( I ) and differentiative conditions ( J ) and treated with 1 mM beta-RA. Data are expressed as mean +- SD. * p < 0.05, *** p < 0.0

Supportive validation

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Experimental details: Fig. 3 m 1 A58 methylation in tRNA regulates PPARdelta translation. a Scheme of tRNA m 1 A-seq. b Comparison of misincorporation signals caused by m 1 A58 in tRNAs between liver CSC and non-CSC cells. Red dots represent tRNAs with elevated m 1 A58 methylation levels in CSCs. c Comparison of translation efficiency (TE) between liver CSC and non-CSC cells. Red dots represent genes with increased TE, while black dots represent genes with decreased TE in liver CSCs. d Frequency of codons corresponding to m 1 A58 elevated tRNAs was higher in the genes with increased translation efficiency (TE) in CSCs. e Unbiased pathway analysis of TE upregulated genes in liver CSCs compared with non-CSCs. f Western blotting analysis of PPARdelta expression in indicated CSCs. n = 4 biologically independent samples. g Effect of elevated m 1 A58 methylation levels in tRNA Ala(AGC) and tRNA Glu(CTC) on protein synthesis. Reporter vectors were transfected into Mock overexpressed PLC/PRF/5 cells, WT-TRMT6/TRMT61A or Mut-TRMT6/TRMT61A overexpressed PLC/PRF/5 cells, respectively. Control vectors were also transfected into these three cell lines to normalize translation differences among these cell lines. The expression level of two synonymous codon control Ala-(GCG) and Glu-(GAA) remained unchanged upon TRMT6/TRMT61A expression level manipulation. n = 6 biologically independent samples. Exact P values from left to right: 0.00021, 0.27, 0.45, 0.00086, 0.21, 0.15. h Representative immunohistochemistry ima

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 Trmt6/Trmt61a knockout inhibits mouse cholesterol synthesis through Ppardelta pathway. a Macroscopic appearance of livers from Trmt6 f/+ /Trmt61a f/+ ; Alb-Cre (WT) and Trmt6/Trmt61a DKO ( Trmt6 f/f /Trmt61a f/f ; Alb-Cre) mice under DEN-induced cancer or in Myc OE mouse model of HCC. Black asterisks indicate liver tumors. Scale bar, 1 cm. b Quantitation of liver tumor numbers of Trmt6 KO, Trmt61a KO, and Trmt6/Trmt61a DKO in DEN-treated and in Myc OE mice. Results are shown as means +- SD ( n = 10 biologically independent mice). Exact P values from left to right: 0.0075, 0.041, 0.032, 0.0037, 0.045, 0.014. c Representative liver H&E staining and immunohistochemistry images of m 1 A, Ppardelta, and Hmgcs2 (brown) from Trmt6 f/ + /Trmt61a f/+ (WT) and Trmt6 -/- /Trmt61a -/- (DKO) in Myc OE and DEN-treated mice. Black circle indicates tumor region. Scale bar, 100 mum. n = 3 independent samples. d Levels of cholesterol biogenesis related proteins in 10-week-old male Trmt6/Trmt61a deleted (DKO) in Myc OE mice transduced with AAV vectors encoding GFP control (oeGFP) or Ppardelta (oePpardelta) for 8 days. WT: Trmt6 f/+ /Trmt61a f/+ ; Myc OE mice. n = 5 biologically independent mice. e Total serum cholesterol levels in 10-week-old normal-fed male Trmt6/Trmt61a deleted Myc OE (DKO) mice transduced with AAV vectors encoding GFP control (oeGFP) or Ppardelta (oePpardelta) for 8 days. WT: Trmt6 f/+ /Trmt61a f/+ ; Myc OE mice. Data are means +- SD. n = 5 mice per group. Exact P val

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 7 TRMT6/TRMT61A complex inhibitor thiram suppresses self-renewal of liver CSCs and tumor growth. a Quantitative LC-MS/MS analysis of m 1 A/A in total RNA and tRNA purified from PLC/PRF/5 cells treated with vehicle or drugs at IC50. Statistical results as means +- SD, n = 5 biologically independent samples. Exact P values from left to right: 0.0048, 0.0086, 0.0037, 0.0090. b Oncosphere forming assays of HCC primary cells following thiram or vehicle treatment. Scale bar, 100 mum. Right panel represents statistical results as means +- SD. n = 15 independent experiments. c Representative magnetic resonance imaging (MRI) images of independent experimental cohorts, of day 0 and day 15 of mice enrolled in treatment with vehicle (DMSO), thiram (1.6 mg kg -1 ), GSK3787 (5 mg kg -1 ) or thiram combined with GSK3787. Red circles and asterisk indicate visible tumor area that was used to calculate tumor volumes. Scale bar, 1 cm. Tumor volumes were calculated on the basis of MRI images from mice bearing liver tumor with a matched initial tumor volume (Right panel). Graph shows means +- SD. n = 6 mice per group. Exact P values from left to right: 0.00065, 0.028. d Tumor volumes were calculated from mice treated with vehicle or atorvastatin (Atorva., 10 mg kg -1 ). Graph shows means +- SD. n = 6 mice per group. Exact P value: 0.0073. e Survival curve derived from mice bearing HCC PDC tumors, treated with vehicle ( n = 11), thiram ( n = 12), GSK3787 ( n = 10), or thiram combined with GSK