Fig. 1
From: Basal p53 expression is indispensable for mesenchymal stem cell integrity
Basal p53 expression maintains MSC integrity. a Quantification of stimulus-induced adipogenic (left panel) and osteogenic (right panel) differentiation of p53+/+ vs. p53−/− MSCs. Data are mean ± S.D. of experiments (n = 4) repeated twice. **p < 0.01, ***p < 0.005 by Student’s t-test. b CFU-F activity of p53+/+ vs. p53−/− MSCs. Data are mean ± S.D. of experiments (n = 6) repeated twice. ***p < 0.005 by Student’s t-test. c Flow cytometric analysis of PI-stained cells showing percentage of p53+/+ vs. p53−/− MSCs in G1, S, and G2 phases of cell cycle. d CAFC activity of p53+/+ vs. p53−/− MSCs. Data are mean ± S.D. of experiments (n = 6) repeated twice. ***p < 0.005 by Student’s t-test. e Methylcellulose colony-forming assays using conditioned media from p53+/+ vs. p53−/− MSCs. Data are mean ± S.D. of experiments (n = 3) repeated twice. *p < 0.05, **p < 0.01, ***p < 0.005 by Student’s t-test. f qPCR analyses of the indicated mRNAs in p53+/+ vs. p53−/− MSCs. All cytokine levels were significantly (**p < 0.01) different between cell types by Student’s t-test (n = 3) except where indicated. g Luminex-based analyses of conditioned media collected from monolayer cultures of p53+/+ vs. p53−/− MSCs. ***p < 0.005 by Student’s t-test. h ChIP showing pull-down of Kitl promoter sequences using a RNA Pol II (positive control) or p53 antibody from cell extracts of p53+/+ MSCs treated with Nutlin-3a (20 µM). **p < 0.01 compared to IgG control by ANOVA and Tukey post hoc test. i p53 mRNA and protein expression quantified by qPCR (upper panel) and immunoblot analysis (bottom panel), respectively, in p53+/+ MSCs transfected with a scrambled (Scr) and p53-specific siRNA. qPCR data are mean ± SD of experiments (n = 3) repeated twice. ***p < 0.005 by Student’s t-test. j, k Effect of siRNAs on CFU-F (j) and CAFC (k) activity in p53+/+ MSCs. Data are mean ± SD of experiments (n = 6) repeated twice. ***p < 0.005 by Student’s t-test. l Quantification of stimulus-induced adipogenic (left panel) and osteogenic (right panel) differentiation of siRNA-transfected p53+/+ MSCs. Data are mean ± SD (n = 4). **p < 0.01 compared to unstimulated control and ++p < 0.01 compared to scrambled (Scr) by ANOVA and Tukey post hoc test
