Fig. 6: SERCA2a associates with autophagy protein ATG5. | Communications Biology

Fig. 6: SERCA2a associates with autophagy protein ATG5.

From: The gut microbiota metabolite trimethylamine N-oxide promotes cardiac hypertrophy by activating the autophagic degradation of SERCA2a

Fig. 6

a-b H9c2 cells transfected with Flag-SERCA2a were cultured in the presence or absence of TMAO (1 mM) for 48 h. Representative images of immunofluorescence detection of ATG5 (red) and SERCA2a (green) in the presence or absence of TMAO. Western blot analysis of ATG5, SERCA2a, LC3, ATG7 and p62 after 48 h of TMAO treatment. c–e Western blot analysis of ATG5 to examine knockdown efficacy. Immunoprecipitation with anti-Flag antibody was performed, n = 3. f–m Western blot analysis (n = 3) of SERCA2a, ANP, and MYH7 and representative microscopic images of cells with rhodamine-phalloidin staining of the cytoskeleton and DAPI staining of the nucleus and cell surface area analysis (n = 4) in H9c2 transfected with control or ATG5 siRNA(100 nM), followed by treatment with TMAO (1 mM) for another 48 h. Scale bar is 40 μm.Two loading controls and target protein have been run on separate gels, and we have indicated it in the supplementary information. Statistical analyse was performed using the One-way ANOVA with Tukey’s multiple comparisons test. Error bars represent S.E.M.

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