Figure 6
From: PML is required for telomere stability in non-neoplastic human cells
PML depletion causes nuclear and chromosomal abnormalities in human fibroblasts. (a) Microphotographs at the indicated magnification of MGG-stained slides of MRC5 cells, transfected with a control non-specific siRNA (CTRL) or PML-KD1 siRNA (PML-KD) for PML RNA interference. (b) Percentage of binucleated cells counted in wild type (Wt), control (CTRL) and PML KD MRC5 cells relative to total asynchronous events evaluated by DAPI staining. A representative experiment of three is shown. One hundred cells were analyzed in each experiment. (c) DAPI nuclear staining (DAPI) and immunofluorescence with anti-PML antibodies (PML) on control (CTRL) or PML KD MRC5 cells, treated with blebbistatin to block cytokinesis. Representative images of NPBs, MNs and nuclear buds (Bs) in PML-depleted cells. (d) Quantitative analysis of overall nuclear abnormalities in PML-depleted cells. NPBs, MNs and Bs were counted and percentages calculated with respect to total cells. A representative experiment of three is shown. One hundred binucleated cells were analyzed in each experiment. (e) Metaphase spreads of control (CTRL) and PML-depleted (PML KD) MRC-5 cells, as above. Examples of chromosomal aberrations are enlarged on the right. (f) Chromosome aberration analysis in control (CTRL) and PML-depleted (PML KD) cells, as above. Bars represent metaphases with at least one aberration events as a percentage of all the scored metaphases. The error bars indicate the s.d. of the counts obtained in three experiments where 50 metaphases were analyzed. The P-value was obtained by a two-tailed Student’s t-test.
