FIGURE 3.
VCP is not involved in CHIKV binding or entry. (A,B) Two hundred thousand HEK 293T cells per well were infected at moi 10 for 1 h in DMEM media or in media with 2 μM CB-5083 on ice or at 37°C, respectively. Cells were then washed twice in PBS, collected, and RNA was extracted. (C) Two hundred thousand HEK293T cells per well were infected at moi 10 for 2 h in DMEM media or in media with 0.5 μM OLX. After 2 h, inoculum was removed, cells were washed twice and incubated for 4 h in media with CB-5083 at 2 μM or media with OLX (0.5 μM) and CB-5083 (2 μM) or media with OLX (0.5 μM), the cells were then washed twice, collected, and RNA was extracted. Relative CHIKV RNA fold change compared to control using 2−ΔΔCT method. Each symbol represents an independent well, data presented are from three independent experiments for panels A and C and two independent experiments for panel B represented as mean ± SD and were analyzed by Mann–Whitney non-parametric two-tailed test for panel D; ∗∗∗p < 0.001.