a) Time course of HIV-1 RT measured by qPCR of early or late RT products at 24 or 48h post-infection. Hela/R5 expressing GFP, CD81GFP or CD81∆cytGFP were infected with HIV-1 (BaL) or HIV-VSV-G. Data are mean fold change ± SEM from 4 independent experiments performed in triplicate, and analysed by two-way ANOVA with Bonferroni’s post-test. b) Hela/R5 cells expressing GFP, CD81GFP or CD81∆cytGFP were infected with single-cycle HIV-1-R5-Luc or HIV-VSV-G-Luc. Data are mean fold-induction ± SEM of luciferase activity from 3 independent experiments performed in triplicate, and analysed by one-way ANOVA with Tukey’s post-test. c) Hela/R5 cells transfected with CRISPR/Cas9-CD81 or left untreated were infected and analysed as in b. Data are mean fold change ± SEM from 3 independent experiments performed in triplicate, and analysed by paired Student t-test, *** p<0.0001 (left) and * p = 0.0157 (right). Left immunoblots show whole cell lysates probed for CD81, and p150 as loading control. d) Hela/R5 cells pre-incubated for 5 days with 2μM cytopermeable peptides containing the CD81 C-terminal sequence (CD81pept) or a scrambled version were infected and analysed as in a.