Validation of microarray data. (*, P < 0.05; **, P < 0.01; ***, P < 0.001.)
A). qRT-PCR showing the effects of MM on ROR1, HDAC4, p53, p21, PMAIP1 and PRDM1 in MES1 and MES7 cells harvested immediately after 24 hour exposure, as well as corresponding xenografts harvested 3 days following the last IP saline or MM injection.
B) Immunoblot analysis (left panel) showing the dose-dependent effects of MM for 24 hours on SP1, ROR1, HDAC4, EZH2, p53, p21, PMAIP1, PRDM1 protein levels in cultured MPM cells; representative immunoblot analysis (right panel) demonstrating the effects of MM on SP1, ROR1, HDAC4, EZH2, p53, p21, PMAIP1, PRDM1 protein levels in subcutaneous MPM xenografts from MM-treated and control mice. Three tumors (T1, T2 and T3) were selected from saline control vs MM 1mg/kg. Tumors were harvested 3 days following the last IP saline or MM injection.
C). Quantitative ChIP analysis of various gene promoters in MES1 cells cultured in normal media with or without MM (100nM, 24 hours).