pSMC1 is localized to nuclear foci in HPV positive cells that become more numerous upon differentiation.
A) Immunofluorescence analysis of undifferentiated HFKs and CIN612 cells for pSMC1 and total SMC1 demonstrates localization to the nucleus in HPV positive cells. Green indicates either pSMC1 (s957) or total SMC1 as indicated. Blue represents nuclear DAPI staining. B). Immunofluorescence of undifferentiated and 72-hour calcium-induced differentiated CIN612 cells identifies large discrete foci of pSMC1 in HPV positive cells. The left panel represents undifferentiated cells while the right panel represents differentiated cells. Green identifies either pSMC1 (s957) or total SMC1 as indicated. Blue represents nuclear DAPI staining. C). The number of pSMC1 foci greater than 3 per nucleus increases upon differentiation of HPV 31 positive cells. The number of pSMC1 nuclear foci in 559 individual undifferentiated and 569 differentiated cells obtained from 3 separate experiments was determined. The graph demonstrates the percentage of total nuclei with greater than 3 foci per nucleus increasing to 80% of cells upon differentiation in high calcium media for 72 hours. A standard students t-test was used to determine statistical significance of <.0001.