Characterization of UHRF1-depleted HeLa cell lines. (A) Stable depletion of UHRF1 by shRNA in HeLa cells as shown by Western analysis (UHRF1 shRNA clones 1-3) compared to clones expressing a nonspecific shRNA (control shRNA clones 1-3). Tubulin levels are shown as loading controls. (B) Representative indirect immunofluorescence of H3K9me3 and H4K20me3 in control and UHRF1 shRNA cells at 40 × magnification. (C-D) Representative indirect immunofluorescence deconvolution of H3K9me3 and H4K20me3 in two UHRF1 shRNA and two control shRNA cell lines. Line traces are shown on the right (blue = DAPI, red = H3K9me3 or H3K20me3) for representative photomicrographs labeled with Greek characters.