Figure 5. The effect of histone modification and DNA methylation enzyme inhibitors on levels of corresponding modifications and gene transcription in HEK293 cells. (A) MeDIP analysis of 5mC levels at Lamc2 gene promoter in DNA purified from cells treated with DZNep (5µM), GSK126 (2.5 µM), or DAC (5µM) for 48 hrs. Mean +SD, n=2 independent experiments, *p<0.05 compared to control. (B) Western blot analysis. Cells were treated with corresponding inhibitors for 48 hrs, and histones were extracted from nuclear pellets. Results of representative Western blot analysis with antibodies to H3K27m3, upper panel, and to histone H3, lower panel, are shown. (C) RT PCR analysis of Lamc2 and control mRNA levels in control cells (CON) and cells treated with DZNep (5µM), GSK126 (2.5 µM), or DAC (5µM) for 48 hrs. Mean +SD, n=4 independent experiments, *p<0.05, ***p<0.0005 compared to control.