FIG. 2.

Latency reactivation for LGIT virus mutants and HIV-1 subtypes in the Jurkat cell system. (A) Infection and serial FACS sorting were performed to isolate the infected, off populations for variants of the LGIT virus. These include mutants of subtype B (mutI Sp1 [S1], mutII Sp1 [S2], mutIII Sp1 [S3], mutI NF-κB [N1]), and mutII NF-κB ([N2]) or variants with U3 regions isolated from the following subtypes: B, A2, A, A/G, B/C, C′, C, B/F, D, F, and H (as in Fig. 1B). One day after FACS sorting (day 22 postinfection [Fig. 1A, panels 5b and 6b]), polyclonal off sorts for WT LGIT mutant and HIV-1 subtype variants were treated with the following pharmacological agents to reactivate latent infections: NF-κB/PKC activators TNF-α (white bars), PMA (gray bars), or prostratin (black bars). Data represent the averages of three independent measurements for each drug perturbation, and error bars are standard deviations. For the LGIT mutants of subtype B, upward and downward arrowheads indicate statistically significant deviations from the wild-type subtype B LTR configuration of LGIT (P < 0.05). The broken gray line at 50% reactivation is drawn as a reference marker. (B) Same as in panel A for latency reactivation by TSA (white bars), SAHA (light gray bars), valproic acid (dark gray bars), or HMBA (black bars). (C) Same as in panel A for latency reactivation using the combination of prostratin and SAHA. Asterisks denote statistical synergism by the combination of drugs relative to the reactivation by either individual agent. For details on the quantitative treatment of synergy, see Materials and Methods.