Figure 5.

Compounds that inhibit spontaneous latency reversal also inhibit CDK9 and Tat-dependent transactivation. (A) CDK9 kinase activity in the presence of compounds, as measured using the ADP-Glo kinase assay. Data are presented relative to CDK9 activity in the presence of 0.1% DMSO. (B) Representative flow cytometry data of GFP expression in CEM-GXR cells transfected with an empty plasmid vector (pCMV-ΔTat) or constitutive Tat expression vector (pCMV-Tat) in the absence or presence of 10 μM apigenin or L7G. (C) Effects of compounds after 24 hours’ incubation on Tat-induced GFP expression in CEM-GXR cells following 24 hours’ incubation. Data are presented relative to Tat-transfected cells treated with 0.1% DMSO. (D) Representative flow cytometry data of mCherry expression and Dendra fluorescence in JurTat cells during no treatment, Dox treatment, or Dox plus 15 μM apigenin. (E) Effect of select compounds (as denoted by the same colors and shapes used in C) on Tat-transactivated mCherry expression in JurTat cells. Data are presented as the ratio of mCherry:Dendra fluorescence and shown relative to the mCherry:Dendra fluorescence in cells treated with 0.1% DMSO. Horizontal dotted lines in A, B, and E denote activities at 100 and 50% of cells treated with 0.1% DMSO.