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. 2019 Mar 1;47(8):3937–3956. doi: 10.1093/nar/gkz128

Figure 3.

Figure 3.

Effects of POLR3G RNAi and ML-60218 on DR2 Alu. (A) Mean relative changes in normalized expression of DR2 Alu RNA in PC-3 cells harvested 48 hrs after transfection with control siRNA or siRNAs targeting POLR3G or POLR3GL, as determined by RT-qPCR in three independent experiments. (B) Mean relative changes in normalized expression of NANOG mRNA in PC-3 cells harvested 48 h after transfection with control siRNA or siRNAs targeting POLR3G or POLR3GL, as determined by RT-qPCR in three independent experiments. (C) Mean relative changes in normalized expression of NANOG-Alu-Sx RNA in PC-3 cells harvested 48 h after transfection with control siRNA or siRNAs targeting POLR3G or POLR3GL, as determined by RT-qPCR in three independent experiments. (D) Mean relative changes in normalized expression of NANOG-Alu-Sx RNA in PC-3 cells harvested 48 h after treatment with or without 20 μM ML-60218, as determined by RT-qPCR in three independent experiments. (E) Mean relative changes in normalized expression of NANOG-Alu-Sx RNA in PC-3 cells harvested 48 h after treatment with or without 20μM ML-60218, as determined by RT-qPCR in 3 independent experiments. Black bars indicate samples that were exposed to 10μg/ml α-amanitin for 4 h prior to harvesting. (F) Mean ratios relative to input of NANOG-Alu-Sx DNA immunoprecipitated using antibodies against POLR3A, POLR2A, H3K79me2 and TAFI48 (negative control) with chromatin from PC-3 cells harvested 48 h after treatment with or without 20 μM ML-60218, as determined by ChIP-qPCR in six independent experiments. (G) Mean ratios relative to input of gene desert DNA immunoprecipitated using antibodies against POLR3A, POLR2A, H3K79me2 and TAFI48 (negative control) with chromatin from PC-3 cells harvested 48 h after treatment with or without 20 μM ML-60218, as determined by ChIP-qPCR in 6 independent experiments. * indicates P < 0.05 relative to control by t-test; ** indicates P < 0.01; *** indicates P < 0.005. # indicates P < 0.05 by t-test for +α-amanitin relative to –α-amanitin. Error bars represent S.E.M.