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. 2014 Oct 15;28(20):2261–2275. doi: 10.1101/gad.250449.114

Figure 5.

Figure 5.

Ssu72 promotes Tat transactivation and regulates S5P-CTD levels at the HIV-1 promoter in vivo. (A) Luciferase activities were measured in extracts of HIV-1:Luc HeLa cells transfected with different siRNAs against Ssu72, Cyclin T1, or AFF1 in the presence or absence of Tat101. The efficiency of siRNA-mediated knockdown was monitored by immunoblot. (B) HIV-1:Luc HeLa cells were transfected with siRNAs specific for Ssu72, RPAP2, or SCP1 in the presence or absence of Tat, as indicated. Luciferase assays were performed as in A. (C) HIV-1:Luc HeLa cells were first transfected with control or Ssu72 siRNAs for 24 h and then were transfected with 5 µg of EGFP or EGFP-Tat101 for another 24 h. The cell extracts were subjected to ChIP analysis with the indicated antibodies. Error bars represent the standard deviation obtained from three independent experiments. Protein lysates were monitored by immunoblot with the indicated antibodies.