Figure 4. Sequential 5‐AzadC + HDACI treatments synergistically activate HIV‐1 gene expression and production in latently infected cells.

• A–F
  J‐Lat 8.4 (A, C, E) and 15.4 (B, D, F) cell lines were mock‐treated or treated with 5‐AzadC. At 48 h post‐treatment, HDACIs were then added for 24 h. At 72 h 5‐AzadC post‐treatment, samples were harvested and analyzed as follows: viral p24 production in the cell supernatant was measured (A, B); FACS analyses were performed and the percentages of GFP ⁺ cells are presented as histograms (C, D); initiated (primers TAR) or elongated (primers tat) transcripts were quantified by RT–qPCR, and results were normalized using the β‐actin gene primers and are presented as histograms representing fold inductions compared to the mock‐treated condition (E, F). Means and standard errors of the means from three independent biological duplicates (n = 6) are indicated. The result obtained with mock‐treated cells was arbitrarily set at a value of 1 (E, F).