Extended Data Fig. 4: GS-CA1 inhibits HIV-1 replication after virus entry and reverse transcription.

a, Quantitative BlaM-Vpr reporter assay for HIV-1 entry. PBMCs were infected with BlaM-Vpr/HIV-1 (NL4-3 strain) in the presence of GS-CA1 or the designated control compounds, loaded with CCF2 substrate dye, and CD3⁺CD4⁺CD8^– T cells containing virus were quantified by flow cytometry to detect CCF2 dye cleavage (indicative of virus entry) after 16 h of incubation. Center line and error bars represent mean ± s.d. values obtained from duplicate cell cultures in each of three independent PBMC donors from a single experiment (n = 3 per group). Significant P values relative to mock-treated HIV-infected samples by unpaired two-tailed Student’s t tests with Welch’s correction are indicated. b, Representative time-of-addition study. MT-2 cells were infected with HIV-1 reporter virus and drugs RPV (93 nM, RT inhibitor), DTG (193 nM, IN inhibitor) and GS-CA1 (30 nM) were added at the indicated time points. Infectivity was measured using a luciferase readout 48 h.p.i. and normalized to mock-treated (DMSO) control. Center line and error bars represent mean ± s.d. values obtained from eight replicate cell cultures per time point and condition from two independent experiments with similar results.