Supplementary Figure 5: Regulation of the phosphorylation level of CTD S5 in vivo. Related to Figure 6.
From: RPRD1A and RPRD1B are human RNA polymerase II C-terminal domain scaffolds for Ser5 dephosphorylation
(a) Chromatin immunoprecipitation (ChIP) experiments using the indicated antibodies showing the effects on S5P levels at various promoters by shRNA-mediated knock-down of RPAP2 in HEK293 cells. *: p < 0.05 compared with shGFP (two-tailed Student’s t-test). Error bars: s.d. from three biological replicates. (b) Western blotting using the indicated antibodies showing the effects on the global S5P and S2P levels by shRNA-mediated knock-down of RPAP2 in HEK293 cells. WT: no lentiviral shRNA infected cells. (c) ChIP experiments using the indicated antibodies showing the effects on S5P levels at various promoters or near promoter regions by siRNA-mediated knocking down RPRD1A in HeLa cell extracts. ACTB (β-ACTIN ): +300 bp region. The S5P levels were normalized to those of RNAPII (detected by N20 antibody as in Fig. 5e). *: p < 0.05 compared with no siRNA transfection control (two-tailed Student’s t-test). Error bars: s.d. from three biological replicates. The bar for β -ACTIN indicates the range of two biological replicates.
