1. A B C Name Sequence TIMP3 promoter
5’/56-FAM/CAGCGGCAA/ZEN/TGACCCCTTGG/31ABkFQ/3’
DBP
5’/56-FAM/CTCTCGGTA/ZEN/TTTTGGCTTCCTGCTTCTC/31ABkFQ/3’ C ZT 24 36 48
si-BMAL1 - +
BMAL1
TIMP3
GAPDH
Supplemental Figure 8. ChIP assay for the promoters of putative CLOCK-regulated genes. (A) The cells at ZT 20 were used for ChIP assay and the promoter sequences of TIMP3 or DBP were analyzed by qRT-PCR after immunoprecipitation with anti-CLOCK antibody or rabbit IgG. The data are presented as the mean ± SD of three independent experiments. (B) The promoter sequences of TIMP3 or DBP are listed. (C) The cells were transfected with siRNAs against BMAL1 and harvested at the indicated ZT time points. BMAL1 and TIMP3 were analyzed by western blotting using each antibody. GAPDH was used a loading control. Two independent western blot analyses were perforemd and representative images are shown.