Fig. 4.

RA caused reduced levels of H3K27me3 at Hox genes in mouse ESCs. a Mouse ESCs were harvested after RA (1 μM) treatment for 24 h, and analyzed by western blotting. Numbers at the bottom were generated by quantification (ImageJ) of the H3K27me3/2/1 signal normalized to the H3 signal. b Immunostaining for H3K27me3 in RA-treated Hep-G2 cell. Direct immunofluorescence analysis was performed. Images were captured by confocal microscope and the nuclei were stained with DAPI. Scale bar: 35 μm. Data are shown as the mean (SD; n = 5). *P < 0.05. c Hox gene mRNA from mouse ESCs treated with RA was assessed by RT-qPCR. RA 1 μM, 24 h. Actb was used as control. Data are shown as mean (SD; n = 3). *P < 0.05. d ChIP assays of H3K27me3 were performed using mouse ESCs treated with 1 μM RA for 24 h. Mouse IgG was used as control. Enrichment of Hox gene promoters was measured by qPCR