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Fig. 4 SGT1-HSP90 pathway regulated MYL12A stability (A) Necdin-SGT1-MYL12 A/B/MYL9 formed a complex in H9C2 cells. An anti-SGT1 antibody, anti-necdin antibody, or control IgG was used to immunoprecipitate cell lysate from H9C2 cells; immune-complexes were blotted with the indicated antibodies. (B and C) RNA interference of Sgt1 decreased MYL12A in H9C2 cells. Representative immunoblots (B) and quantification of three independent experiments each group (C) are shown. Data are presented as means ± SEM, n = 3; ??p < 0.01, unpaired t test. (D and E) Overexpression of SGT1 increased the MYL12A protein levels in H9C2 cells. Representative immunoblots (D) and quantification of three independent experiments (E) are shown. Data are presented as means ± SEM, n = 3; ??p < 0.01, unpaired t test. (F) RNA interference of Sgt1 increases the ubiquitination level of MYL12A. H9C2 cells were transfected with the indicated plasmids or siRNAs. Cell lysates were immunoprecipitated with an antibody against Myc, and ubiquitination was detected with an anti-HA antibody. (G-J) Representative immunoblots (upper lane) and statistics data of three independent experiments (lower lane) from H9C2 cells following treatment with different concentrations of HSP90 inhibitors 17-AAG (G and H) or geldanamycin (GA) (I and J). Data are presented as means ± SEM, ?p < 0.05; ??p < 0.01, post hoc Dunnett?s t test, one-way ANOVA.