Figure 6 : CRY1 accelerates ubiquitin-mediated FOXO1 degradation.

From: SREBP1c-CRY1 signalling represses hepatic glucose production by promoting FOXO1 degradation during refeeding

Figure 6

(a) Mouse primary hepatocytes were adenovirally infected with Ad-MOCK or Ad-CRY1. The cells were treated with 20 μM MG132 or vehicle for 4 h. Total cell lysates were analysed by western blotting with indicated antibodies. (b) HEK293T cells were transfected with GFP-CRY1 and/or FOXO1-MYC expression vectors. Co-immunoprecipitation with an anti-MYC antibody and western blotting were performed with the indicated antibodies. IP, immunoprecipitation. (c) COS-1 cells were co-transfected with plasmids encoding FOXO1-MYC, GFP-CRY1, and Ubiquitin-HA. After transfection, the cells were treated with MG132 (20 μM) for 6 h and then the cell lysates were subjected to immunoprecipitation with an anti-MYC antibody followed by western blotting with indicated antibodies. IP, immunoprecipitation. (d) Mouse primary hepatocytes were infected with Ad-MOCK or Ad-CRY1. After infection, the cells were treated with MG132 (20 μM) for 4 h. Nuclear and cytosolic fractions were isolated and analysed by western blotting with indicated antibodies. (e) COS-1 cells were co-transfected with plasmids encoding nFOXO1-MYC, GFP-CRY1, and Ubiquitin-HA. After transfection, the cells were challenged with MG132 (20 μM) for 6 h. The cell lysates were subjected to immunoprecipitation with an anti-MYC antibody. IP, immunoprecipitation. See Supplementary Fig. 13 for original full immunoblot.