Research Paper Volume 5, Issue 7 pp 531—538

Figure 3. Favoring an escape from PDGFB-induced senescence increases PDGFB-induced transformation. Fibroblasts were infected with a ctrl, a PDGFB- with or without an shRNA against p53 encoding vector. (A) RNAs were extracted and reverse-transcribed. Quantitative PCRs were performed against PDGFB and the expression normalized with ACTB. (B) Twenty five thousand cells were seeded in a well of 12-well plate, and analyzed for their SA-β-Gal activity 3 days later. (C) Cells were seeded as indicated in (B) and fixed 2 days later with PFA 4%. After DNA staining by Hoechst, percentage of SAHF-positive cells was calculated. (D) Seventy five thousand cells were seeded in a well of 6-well plate and cultured during 7 days. Representative photographs are presented. (E) Indicated cells were seeded at high densities. Two weeks later they were PFA-fixed and stained with crystal violet. (F) Fifty thousand cells were plated in agar in 6-well plates. After 3 weeks colonies were quantified and photographed. All histograms shown in this figure represent the mean±SD. Three different experiments were performed and one representative is shown. t-test was used for statistical analysis. *** indicated p value < 0.005.