MOESM4 of HIV-1 capsid is involved in post-nuclear entry steps

Additional file 4: Fig. S4. Conditions for the CsA washout assay. (A) H126Q cells were infected with WT HIV-1GFP vector by spinoculation in the presence of CsA (1 μM) or C-A1 (3 μM). The drug was washed out at the indicated time points (time of washout) and cells were analysed by FACS 48 h later to determine the percentage of infected (GFP+) cells. (B) Same as (A) but T5Cyp cells expressing functional human TRIMCyp were used. (C) A prolonged time of uncoating assay, in which CsA was washed out at the indicated time points. Rescue of infection reaches a plateau after 10 h. Average ± SD of three independent experiments are shown in (A-C)

MOESM4 of HIV-1 capsid is involved in post-nuclear entry steps

Additional file 4: Fig. S4. Conditions for the CsA washout assay. (A) H126Q cells were infected with WT HIV-1GFP vector by spinoculation in the presence of CsA (1 μM) or C-A1 (3 μM). The drug was washed out at the indicated time points (time of washout) and cells were analysed by FACS 48 h later to determine the percentage of infected (GFP+) cells. (B) Same as (A) but T5Cyp cells expressing functional human TRIMCyp were used. (C) A prolonged time of uncoating assay, in which CsA was washed out at the indicated time points. Rescue of infection reaches a plateau after 10 h. Average ± SD of three independent experiments are shown in (A-C)

MOESM5 of HIV-1 capsid is involved in post-nuclear entry steps

Additional file 5: Fig. S5. CsA washout assays in cells depleted of Nup153 or control DsRed cells. T5Cyp cells expressing an shRNA against Nup153 or DsRed (control) were infected at an MOI of 0.01–0.05 with an HIV-1GFP vector (WT) in the presence of CsA (1 μM). The drug was washed out at the indicated time points (time of washout) and cells were analysed by FACS 48 h later to determine the percentage of infected (GFP+) cells. To control for specificity, cells were infected in the same way using the N74D mutant virus. Raw data of three independent experiments are shown. The data have been used to compile fold rescue levels shown in Figure 6