Figure 2.

HPV16 infectivity is rescued by restoring A2t expression. (a) WT HeLa cells were transiently transfected with a control plasmid and A2t KO cells were transfected with either an mCherry-tagged AnxA2 expression plasmid or a non-target control plasmid. AnxA2 expression was restored 24 h post-transfection, stabilizing S100A10 and restoring heterotetrameric A2t (western blot, lane 3). Lysates were prepared with total populations of transfected cells, and mCherry-AnxA2 transfection efficiency was approximately 50%. Western blot images were cropped. See full-length blots with molecular weight standards in Supplementary Fig. 1b. (b) 24 h post-transfection, WT and A2t KO cells + non-targeting plasmid control and A2t KO cells + an mCherry-AnxA2 expression plasmid were treated with HPV pseudovirions (TCID₅₀) carrying a GFP reporter plasmid. Infection was measured 48 h p.i. via flow cytometry, and infection rescue was measured by gating on mCherry and analyzing the %GFP-positive cells within that population. See mCherry gating strategy in Supplementary Fig. 2. Results show the mean %GFP-positive cells ± s.d. (n = 3, normalized to WT), and are representative of at least 3 independent experiments. Statistics: 1-way ANOVA with Dunnett’s multiple comparisons test – ****P < 0.0001.