MOESM1 of NNRTI-induced HIV-1 protease-mediated cytotoxicity induces rapid death of CD4 T cells during productive infection and latency reversal

Additional file 1. Fig. S1. Reporter viruses employed in this study. Diagram of the main characteristics of the reporter HIV-1 viruses employed in this study (see methods). Figure S2. Stability of GFP and HSA reporter proteins following apoptosis. T cells infected or not with GFP (NLENG1-ES-IRES) or HSA (NLENHSA-ES-IRES) reporter viruses were treated or not with staurosporine (1 μM) at 5 dpi. Twenty-four hours later GFP fluorescence and HSA expression (following anti-HSA staining) were measured by flow cytometry. Figure S3. Toxicity of NNRTI treatment on uninfected cells. Additional data from experiment in Fig. 1c, d here showing effects of NNRTIs on mock infected T cells. Figure S4. EFV killing of Infected resting T cells. Additional data from experiment in Fig. 2a showing killing with EFV. Figure S5. Addition of fresh IL-7 does not prevent NNRTI killing. IL-7 treated resting CD4 T cells infected with HSA reporter were treated at 5 dpi with EFV in the presence of freshly supplemented IL-7 or not. Cell death was analyzed by flow cytometry measuring FSC reduction of HSA+ cells after 24 h. Data are representative of 2 independent experiments. Figure S6. Inhibition of NNRTI killing by PIs IDV, SQV and TPV. T cells infected with NL4-3 virus were treated at 5 dpi with RPV for 4 h in the presence of various concentrations of the PIs IDV, SQV and TPV. Data represent the percentage of inhibition of RPV killing. Productively infected cells were identified by intracellular p24Gag staining. Data are representative of 2 experiments.