Fig. 7: A-485 destabilizes hepatic FOXO1 protein. | Cell Death & Disease

Fig. 7: A-485 destabilizes hepatic FOXO1 protein.

From: Selective inhibition of CBP/p300 HAT by A-485 results in suppression of lipogenesis and hepatic gluconeogenesis

Fig. 7

a Endogenous FOXO1 protein were precipitated in HepG2 cells treated with 3 μM A-485 and 100 μM 8-Br-cAMP for 6 h and acetylation of FOXO1 was detected. b Cytoplasmic (cyto) and nuclear (nucl) protein levels of FOXO1 in primary mouse hepatocytes treated with 3 μM A-485 and 100 μM 8-Br-cAMP. c Immunofluorescence analysis of FOXO1 (red) localization in HEK293T cells transfected with plasmid DNA expressing HA-FOXO1 for 24 h and treated with 3 μM A-485 and 100 μM 8-Br-cAMP for 3 h. d, e FOXO1 levels in primary mouse hepatocytes treated with 3 μM A-485 and 10 μg/ml cycloheximide (CHX) for the indicated time in the presence of 100 μM 8-Br-cAMP. Signal intensity was quantified by image J software for statistical comparison. f, g FOXO1 protein level in primary mouse hepatocytes treated with 100 μM 8-Br-cAMP, 3 μM A-485, 10 μg/ml CHX, and 10 μM MG132. Signal intensity was quantified by image J software for statistical comparison. h HepG2 cells co-expressed with HA-tagged FOXO1 and Flag-tagged ubiquitin were treated with 3 μM A-485 for 6 h. FOXO1 ubiquitination level was detected. i The interaction between FOXO1 and COP1 was detected in HepG2 cells treated with 3 μM A-485. Data are expressed as means ± SEM for three independent experiments. **P < 0.01 vs. corresponding control group.

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