459000

Lood C, Blanco LP, Purmalek MM, Carmona-Rivera C, De Ravin SS, Smith CK, Malech HL, Ledbetter JA, Elkon KB, Kaplan MJ
Nature medicine 2016 Feb;22(2):146-53

Assembly of human mitochondrial ATP synthase through two separate intermediates, F1-c-ring and b-e-g complex.

Fujikawa M, Sugawara K, Tanabe T, Yoshida M
FEBS letters 2015 Sep 14;589(19 Pt B):2707-12

Population of ATP synthase molecules in mitochondria is limited by available 6.8-kDa proteolipid protein (MLQ).

Fujikawa M, Ohsakaya S, Sugawara K, Yoshida M
Genes to cells : devoted to molecular & cellular mechanisms 2014 Feb;19(2):153-60

Tau oligomers impair memory and induce synaptic and mitochondrial dysfunction in wild-type mice.

Lasagna-Reeves CA, Castillo-Carranza DL, Sengupta U, Clos AL, Jackson GR, Kayed R
Molecular neurodegeneration 2011 Jun 6;6:39

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-ATP5H Mouse Monoclonal Antibody (Product # 459000) and a 21 kDa band corresponding to an isoform of ATP5H was observed in cell lines tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), Caco-2 (Lane 2), HCT 116 (Lane 3) and Hep G2 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A 25 kDa band (*) corresponding to tissue IgG was observed in mouse tissues.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of ATP5H was performed using Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ATP5H Mouse Monoclonal Antibody (Product # 459000) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the composite image showing mitochondrial pattern of ATP5H. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

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

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Tau oligomers induce mitochondrial alterations . (A) Representative western blot of mouse brain homogenate. The levels of complex I, complex V, and caspase-9 were measured by band quantification and normalized with the levels of tubulin. Abbreviations are as in Fig. 4. (B) Complex I levels were significantly lower in the hemisphere injected with tau oligomers in comparison with the ones injected with fibrils or monomers. (C) No differences in the levels of complex V were observed in any of the groups. (D) Caspase-9 activation was significantly higher in tau oligomer groups over both tau fibril- and monomer-injected groups. (E-G) Double staining between human tau-specific antibody HT7 (green fluorescence) and mitochondrial porin antibody (red fluorescence) demonstrates the internalization of tau oligomers in CA1 cells and their interaction with the mitochondria. (H) The co-localization of tau oligomers with mitochondria was confirmed by ICA of the signal. Data are represented as mean +- SE. *p < 0.01, n = 6.