Fig. 3.

Latency-reversal assay testing analogs of the thioredoxin reductase inhibitor pleurotine. According to literature, pleurotine, a compound showing high structural similarity to NSC 401005 (D75), irreversibly inhibits TrxR, which is one of the two key components in the Trx/TrxR redox system. Three compounds that inhibit the Trx/TrxR system (auranofin, PX12, and tiopronin) were tested at different concentrations for their latency-promoting potential in combination with TNF-α (10 ng/mL). PI staining was introduced to gauge cytotoxicity of the inhibitors. (A) Reactivation percentage from the experiment. Auranofin at 2 μM increased HIV latent reactivation. NSC 401005, PX12, and tiopronin showed suppression effects at different strengths, with NSC 401005 at 10 μM and PX12 at both concentrations showing the strongest effects. (B) Live percentage from the experiment, measured by the proportion of cells that are PI-negative among all cells measured (see SI Appendix, Fig. S7 for gating). Auranofin at 2 μM and NSC 401005 at 5 μM did not reduce cell viability. NSC 401005 at 10 μM and PX12 at 60 and 80 μM caused notable cell death. Tiopronin kept cells alive at both concentrations in both JLat clones. Significant P values are labeled with asterisks (* for familywise error rate ≤ 0.1, ** for familywise error rate ≤ 0.05). See Materials and Methods for details.