FIG 5 .

Distinct populations of foci exist during HPV infection. (A and B) CIN612 cells were differentiated for 72 h in 1.5 mM calcium medium. Immunofluorescence analysis was performed on cells stained with anti-FANCD2 (green) and either anti-BRCA1 or anti-p-SMC1 (red). Cells were then counterstained with anti-γH2AX (pink) and DAPI (blue). Arrows indicate foci where FANCD2, γH2AX, and p-SMC1 are found together. UD, undifferentiated; D, differentiated. (C) The graph demonstrates the percentage of cells with FANCD2 foci where at least one focus colocalizes with γH2AX and either BRCA1 or p-SMC1. (D) The graph represents the percentage of all HPV-positive cells where at least one FANCD2 focus colocalizes with γH2AX and either BRCA1 or p-SMC1. Error bars represent the standard deviations between experiments. A standard Student’s t test was used to determine statistical significance. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001. ns, not significant. (E) Representative image of three distinct populations in differentiated CIN612 cells stained with anti-FANCD2 (green), anti-p-SMC1 (red), anti-γH2AX (pink), and DAPI (blue). Populations are identified as having FANCD2 foci with no p-SMC1 foci (i), having p-SMC1 foci with no FANCD2 foci (ii), and having both FANCD2 and p-SMC1 foci (iii).