Fig. 1
HDAC4 is dysregulated by UPS-mediated degradation during different models of senescence and aging. a Immunoblot analysis in IMR90 cells undergoing replicative senescence, using the indicated antibodies. The percentage of SA-β-gal positive cells is indicated. Actin was used as loading control. b Representative microscopic images of SA-β-gal stained IMR90 cells (scale bar 50 μm). c Immunoblot analysis in tissue-derived lysates obtained from C57BL/6 J female mice sacrificed at 128 (young) and 774 (old) days of age using the indicated antibodies. Actin was used as loading control. d Immunoblot analysis in BJ/hTERT cells expressing the different transgenes for the indicated time. Vimentin was the loading control. The percentage of SA-β-gal positive cells is indicated. e Cellular lysates were obtained from BJ/hTERT cells expressing the indicated transgenes. Immunoblots were performed using the indicated antibodies. The percentage of BrdU positive cells is indicated. f Cellular lysates were obtained from BJ/hTERT cells expressing AKT1 or Hygro as control. Immunoblots were performed using the indicated antibodies. The percentage of SA-β-gal positive cells is indicated. g Cellular lysates were obtained from IMR90 cells expressing the indicated transgenes. Immunoblots were performed using the indicated antibodies. The percentage of SA-β-gal positive cells is indicated. h Immunoblot analysis of HDAC4, HDAC5, and p62 in BJ/hTERT and BJ/hTERT/RAS expressing cells after 12 days of culture and 8 h of treatment with MG132 (1 μM) and chloroquine (10 μM), as indicated. SMC3 was used as loading control. i Cellular lysates obtained in BJ/hTERT expressing for 8 days the indicated transgenes and treated or not for 8 h with MG132 were immunoprecipitated using anti-HDAC4 and immunoblotted with the indicated antibodies. j Immunoblot analysis using the indicated antibodies in BJ/hTERT cells expressing HRASG12V and silenced or not for GSK3β. RAS expression was induced by DOX treatment