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. 2018 Jan 8;25(4):677–690. doi: 10.1038/s41418-017-0004-4

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© ADMC Associazione Differenziamento e Morte Cellulare 2018

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License, which permits any non-commercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. If you remix, transform, or build upon this article or a part thereof, you must distribute your contributions under the same license as the original. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/.

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Fig. 4 — TWIST2 expression is downregulated in MSCs following p53 inactivation. a qPCR (top panel) and immunoblot (bottom panel) analyses of TWIST2 mRNA and protein levels, respectively, in p53^−/⁻ vs, p53^+/+ MSCs. qPCR data are mean ± S.D. of experiments (N = 2) repeated twice. ***p < 0.005 by Student’s t-test. b ChIP showing pull-down of Twist2 promoter sequences using a RNA Pol II (positive control) or p53 antibody from cell extracts of MSCs treated with Nutlin-3a (20 µM). *p < 0.05, **p < 0.01 compared to IgG control by ANOVA and Tukey post hoc test. c Effect of siRNAs on Twist2 mRNA levels as determined by qPCR. Data are mean ± S.D. of experiments (N = 2) repeated three times. ***p < 0.005 by Student’s t-test. d qPCR (top panel) and immunoblot (bottom panel) analyses showing effect of siRNAs on TWIST2 mRNA and protein expression, respectively, in MSCs. qPCR data are mean ± S.D. of experiments (N = 2) repeated twice. **p < 0.01 compared to Scr by Student’s t-test. e Densitometry (top panel) and immunoblot (lower panel) showing effect of siRNAs on BAX and p53 protein expression in MSCs. f, g Effect of siRNAs on CFU-F (f) and CAFC activity (g) in MSCs. Data are mean ± S.D. of experiments (N = 6) repeated twice for CAFC and three times for CFU-F. ***p < 0.005 compared to Scr by Student’s t-test. h Effect of siRNAs on stimulus-driven adipogenic (left panel) and osteogenic (right panel) differentiation. Data are mean ± S.D. of experiments (N = 4). **p < 0.01 compared to CCM, ⁺⁺p < 0.01 compared to Scr + AIM or Scr + OIM by ANOVA and Tukey post hoc test