Fig. 2.

H2Bub1 regulates a group of mitochondrial respiratory genes. a Microarray analysis was previously reported in HEK293T cells transfected with Myc-tagged H2BK120R plasmid or with empty Myc plasmid (Cont.) to predict the downstream targets of H2Bub1 [38]. A group of genes encoding the mitochondrial respiratory chain were reported to be affected following H2BK120R overexpression [38]. The affected genes are listed. b, c Experimental validation (RT-PCR analysis) of the H2Bub1-affected genes listed in a. H1299 cells transfected with GFP-tagged wild-type H2B (H2BWT-GFP) or K120R-mutated H2B (H2BK120R-GFP) and control siRNA (siCont.) or RNF20-specific siRNA (siRNF20) were lysed and subjected to RT-PCR analysis. d Western Blot analysis of the selected H2Bub1-affected genes and the related quantitative analysis of protein expression levels. H1299 or A549 cells transfected with control siRNA (siCont.) or RNF20-specific siRNA (siRNF20) were lysed and subjected to western blot analysis with antibodies as indicated. e Chromatin immunoprecipitation (Ch-IP) analysis was performed with anti-H2Bub1 antibody or normal mouse IgG (NIgG). Two intergenic regions were used as negative controls. f Chromatin immunoprecipitation (Ch-IP) analysis was performed with anti-Pol-II antibody or normal rabbit IgG (NIgG). An intergenic region was used as a negative control. The results of d are shown the mean with 95% CI, and the others are shown as mean ± S.D. n = 3 per group. *p < 0.05; **p < 0.01; ***p < 0.001