CRTC2 binds Bmal1 promoter.
A, Bmal1 promoter deletion analysis. HEK293T cells were co-transfected with CRTC2, β-gal, and luciferase reporter plasmids driven by different Bmal1 promoter fragments as indicated. Luciferase activity was measured after 4-h forskolin (10 nm) treatment and normalized to β-gal activity. B, ChIP analysis showing the effect of 30-min glucagon treatment on recruitment of CRTC2 to the promoters of Bmal1 and G6Pase in primary hepatocytes (NS: nonspecific binding). IP, immunoprecipitation. C, ChIP analysis showing effect of 4- and 24-h fasting on the recruitment of CRTC2 to the promoters of Bmal1 and G6Pase in mice liver (n = 3 per condition, * p < 0.05, ** p < 0.01, ns, no significance). Ad lib, ad libitum.