FIG 4 .

Comet EdU assays demonstrate that nascent DNA contains minimal breaks upon calcium-induced differentiation. Comet-EdU assays were performed on undifferentiated and calcium-differentiated HPV-positive cells, which were visualized using epifluorescence microscopy. Cells were pulse labeled with EdU followed by neutral comet assays and stained for total DNA and EdU. Green signal indicates total DNA stain, while red signal indicates nascent DNA (EdU). Undifferentiated cells contain a positive signal in the red tail, indicating the presence of breaks in nascent DNA, while differentiated cells contain minimal red signal in the tail, indicating minimal breaks in nascent DNA upon differentiation. Images are representative images from three individual experiments.