Figure 7 : CRY1 is involved in MDM2-mediated FOXO1 ubiquitination.

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

Figure 7

(a) Mouse primary hepatocytes were infected with Ad-MOCK or Ad-CRY1 and/or siCON or siMDM2. Total cell lysates were analysed by western blotting with indicated antibodies. (b) HEK293T cells were transfected with FLAG-MDM2, nFOXO1-MYC, and GFP-CRY1 expression vectors. Total cell lysates were subjected to co-immunoprecipitation with an anti-MYC antibody followed by western blotting with indicated antibodies. IP, immunoprecipitation. (c) COS-1 cells were co-transfected with plasmids encoding nFOXO1-MYC, FLAG-MDM2, FLAG-CRY1, and Ubiquitin-HA. After transfection, the cells were challenged with MG132 (20 μM) for 6 h. Cell lysates were subjected to immunoprecipitation with an anti-MYC antibody. IP, immunoprecipitation (d) COS-1 cells were co-transfected with plasmids encoding nFOXO1-MYC, FLAG-MDM2, Ubiquitin-HA, and siCRY1. Cells were treated with MG132 (20 μM) for 6 h. Cell lysates were subjected to immunoprecipitation with an anti-MYC antibody. IP, immunoprecipitation. See Supplementary Fig. 13 for original full immunoblot.