Substrate Stiffness Influences Doxorubicin-Induced p53 Activation via ROCK2 Expression

Figure 2

Soft substrates diminish doxorubicin-induced p53 activation. (a–c) Cells infected with a control or p53 shRNA-expressing retrovirus were cultured on substrates with elasticities of 2 and 30 kPa. (a) The expression of p21^Waf1, NOXA, and PIG6 in cells cultured in the presence or absence of DOXO (1 μg/mL) for 24 h was evaluated by quantitative real-time PCR. Each bar represents the mean ± SD; . Asterisks, . (b-c) Control and p53 shRNA-expressing cells were treated with DOXO for the indicated time periods. (b) Total cell lysates were subjected to immunoblot analysis with antibodies against p21, Lamin B1, and α-tubulin as a loading control. The white arrowhead indicates full-length Lamin B1, while the black arrowhead indicates a cleaved fragment. (c) Total cell extracts, nuclear extracts, and cytosol extracts were subjected to immunoblot analysis with antibodies against p53, HDAC1 as a nuclear marker, and α-tubulin as a loading control or cytosol marker.

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