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. Author manuscript; available in PMC: 2012 May 20.
Published in final edited form as: Mol Cell. 2011 May 20;42(4):477–488. doi: 10.1016/j.molcel.2011.03.011

Figure 6. TFIIS is recruited to EGF targets, and counters RNF20 in the transcriptional response to EGF.

Figure 6

(A) ChIP analysis of relative TFIIS abundance in HeLa cells stimulated with EGF (20 ng/ml) for 0, 0.5 hours, 1 hour or 2 hours. Prior to EGF addition, cells were serum starved for 16 hours. Immunoprecipitated DNA and input DNA were quantified by qPCR with primers for the EGF target genes RHOB, FOSL2, NR4A2, FOS and TM4SF1, and also for GAPDH and p21, which are not responsive to EGF. Bars indicate averages of data from duplicate qPCR reactions; error bars represent standard deviation. Similar data was obtained in 3 independent experiments.

(B,C) qRT-PCR analysis of total mRNA (B) or heterogeneous nuclear precursor RNA (C) levels of the indicated EGF target genes at various times after exposure to EGF of HeLa cells transiently transfected with TCEA1+TCEA2 siRNA (light blue) or LacZ siRNA (black). All values are normalized to GAPDH mRNA. Precursor RNA levels were measured with primer pairs derived from intronic sequences of the indicated genes. Values indicate averages of data from duplicate qPCR reactions; error bars represent standard deviation. Similar data was obtained in 4 independent experiments. See also Supplemental Fig. S3 for parallel analysis after TCEA1 knockdown alone.