Fig. 8

Timeless is transcriptionally activated by H3K27 acetylation. (a) Genome bioinformatics analysis showed that the Timeless promoter was highly enriched for H3K27ac.(cited from http://genome.ucsc.edu/) (b) Chromatin immunoprecipitation (ChIP) was performed to measure the H3K27ac enrichment in HCT116 cells with high a Timeless level and CX-1 cells with a low Timeless level. Timeless expression was measured by RT-qPCR (c) and protein levels of Timeless, H3K27ac, and total Histone-3 were measured by western blotting (d) in HCT116 and RKO cells treated with C646 or DMSO for 48 h. The enrichment of H3K27ac in the Timeless promoter was assessed via ChIP assay in HCT116 cells silenced by CBP (e) and P300 (f). Timeless, CBP (g) and P300 (h) mRNA expressions were measured by RT-qPCR and protein levels of CBP (i), P300 (j), Timeless, and H3K27ac in HCT116 and RKO cells were measured by western blotting