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. 2021 Mar 10;218(5):e20202033. doi: 10.1084/jem.20202033

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© 2021 Qu et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 2. — MDM4 is a matrix stiffness–regulated mechanosensitive inhibitor of p53. Primary human lung fibroblasts were cultured on 1–20-kPa PA gels. (A) mRNA levels of MDM4 and MDM2 were determined by qPCR. (B) Protein levels of MDM4 and MDM2 were determined by immunoblot. (C) Levels of acetylated p53 (Ac-p53) and total p53 were determined by immunoblot. (D) Human lung fibroblasts were cultured on 1- or 20-kPa PA gels in the presence of 50 nM MDM4 (M4) siRNAs (siR) or control siRNAs. Levels of MDM4, Ac-p53, and total p53 were determined by immunoblot. (E) Human lung fibroblasts infected with MDM4-expressing lentivirus (Lenti-M4) or the control lentivirus (Lenti-Ctrl) were cultured on 1- or 20-kPa PA gels. Levels of MDM4, Ac-p53, and total p53 were determined by immunoblot. GAPDH was used as internal reference control in qPCR analysis and as loading control in immunoblot analysis. Results are the means ± SD of three or four separate experiments. **, P < 0.01 (ANOVA). k, kilodalton.