Figure 5. STRAP affects the local chromatin modification at HES1 and HES5 loci in sphere-derived cells.

A. The fraction of CD133+/CD44+ cells from the adherent and sphere cell population (HCT116, DLD-1, and HT-29) was analyzed by flow cytometry and plotted as % positive cells. *P< 0.05, vs. adherent group. B. Cell lysates from the adherent and sphere cells (HCT116, DLD-1, and SW620) were collected and subjected to immunoblot analyses with the indicated antibodies. C. Five-day old sphere-derived cells were used to establish STRAP knockdown clones and subsequent ChIP assay. Western blot analyzed the expression of STRAP in sphere-derived cells. D, E, and F. Anti-H3K4me3, -H3K27me3, -EZH2, and -SUZ12 antibodies were used for ChIP assays in sphere-derived cells. PCR amplification was done with a series of primers targeting distal, middle, and proximal sequences (see Supplementary Table S1 for primer sequences) in HES1 and HES5 loci. n=3, *P< 0.05, shSTRAP#1 vs. shCtrl in sphere-derived HCT116 and DLD-1 cells.