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. 2020 Aug 13;11:4051. doi: 10.1038/s41467-020-17753-w

Fig. 1. Regulation of host cell survival, but not HIV-1 reverse transcription and integration into the host genome by inhibition of autophagy.

Fig. 1

a CMT transfected with Atg7 siRNA were infected with HIV-1 (NL4-3, 0.1 MOI) and cultured for 4 d to establish latency. After latency reversal by PHA, p24+ cells (n = 3 biologically independent samples) and Atg7 expression (representative of two biologically independent experiments) were determined. *p = 0.011. b CMT latently infected with HIV-1 were stimulated with IDB with SAR405 (2 μm) or CQ (10 μm). p24 staining and the number of LC3 punctate per cells were analyzed. p24+ cells (n = 3 biologically independent experiments), p = 0.010 (SAR405) and 0.0098 (CQ); LC3 punctate/cell (n = 30 cells from two biologically independent experiments), p = 0.0001 (no virus) and 0.0001 (HIV-1). c, d Atg7 siRNA-transfected CMT were infected with HIV-1. Alternatively, CMT infected with HIV-1 were cultured with SAR405. Cells at different time after infection were collected (n = 3 biologically independent samples) for RT-PCR for R/U5c and LTR-gagd. eAlu-gag PCR for genomic DNA from CMT as in a, CMT treated with SAR405 as in b or uninfected CMT. Data were normalized against β-globin. ND: not detectable. Data are presented as mean ± SD (n = 3 biologically independent samples). f, g CMT cultured as in a, b were reactivated with PHA for 24 h f (n = 3 biologically independent samples). PBMCs from ART-treated HIV-1-infected patients (n = 5 patients) were stimulated with IDB in the presence or absence of SAR405 for 24 h g. HIV-1 mRNA was determined by RT-PCR. Data are presented as mean ± SD. The dashed line indicates detection limit. h, i CMT latently infected with HIV-1 (NL4-3, 1 MOI) as in a were reactivated with 100 nm IDB for 24 h. Annexin V h or DEVD i staining were analyzed by flow cytometry (n = 3 biologically independent samples). SAR405 vs. control, p = 0.0012 h, 0.0024 i; CQ vs. control, p = 0.0164 h, 0.0088 i. Source data are provided as a Source Data file.