Fig. 6: Egr-1/BMAL1/CLOCK regulates the robustness and rhythm of Cidea by inhibiting its transcription.

A mRNA expression of Cidea at the indicated time points in livers from C57BL/6 J mice at 2 months, 6 months, and 12 months of age (2 months ZT1: n = 6, ZT5: n = 5, ZT9,13,17,21:n = 4; 6 months ZT1: n = 4, ZT5,13: n = 6, ZT9:n = 5, ZT17,21: n = 4; 12 months: n = 4 biologically independent animals); B Protein levels of Cidea at the indicated time points in livers from B6 mice at 2 months, 6 months, and 12 months of age that were entrained to a 12-h light/dark cycle. C Quantitative analysis of Cidea protein levels in Fig. 6B (n = 4 biologically independent animals per group). D Protein levels of Cidea in primary hepatocytes isolated from 6-month-old C57BL/6 J mice after deletion of Egr-1. E Protein levels of Cidea in primary hepatocytes isolated from 6-month-old C57BL/6 J mice overexpressing Egr-1. F ChIP assay (n = 3 or 4 biologically independent samples per group); G Luciferase assay (n = 5 biologically independent samples per group). H HEK293T cells were co-transfected with Myc-BMAL1, Egr-1, and CLOCK expression plasmids. Co-IP of Egr-1, Myc-BMAL1, and CLOCK was performed. E means Egr-1, B means BMAL1, and C means CLOCK. Each experiment was repeated three times independently. The data represent the mean ± SEM. Exact p values are depicted in the figure. Orange color p value means 6 months versus 2 month group; Purple color p value means 12 months versus 2 month group; Black color p value means 6 months versus 12 month group. Statistical analysis was performed using one-way ANOVA. Source data are provided as a Source Data file.