Figure 3
From: HDAC1 inhibition by MS-275 in mesothelial cells limits cellular invasion and promotes MMT reversal
Effect of HDAC1 siRNA silencing on E-cadherin and TGFβ1 expression in MCs and effect of MS-275 in blocking MMT induced by exposure to PD fluid. (A) Quantitative RT-PCR was performed on total RNA from MCs transfected with either control (siCtrl) or specific HDAC1-targeting siRNAs (siHDAC1). Expression of E-cadherin, TGFβ1, HDAC1 was evaluated. Bars represent means ± SEM of 5 experiments. (B) Western blots showing the expression of E-cadherin and HDAC1 in total cell lysates of MCs from three different PD patients transfected with either control or specific HDAC1-targeting siRNAs. GAPDH expression was evaluated as a loading control (C) Effect of MS-275 on MMT induced by exposure to PD fluid. Quantitative RT-PCR was performed on total RNA from epithelial-like MCs treated for seven days with stay safe balance 4.25% (used at a concentration of 1:1 with complete 199 Medium). Alternatively, cells were treated for four days with stay safe balance 4.25% (used at the same concentration as before) and then treated with the same PD fluid in the presence of MS-275 for three more days. As a control, MCs were left untreated for four days and then treated with MS-275 or DMSO for the last three days. Expression of MMP2, Snail, PAI-1, TGFβ1, TGFβRI, Occludin was evaluated on total RNA by qRT-PCR. Bars represent means ± SEM of 4 experiments (D) Western blots showing the expression of E-cadherin, Occludin and Snail in total cell lysates of MCs treated as above. GAPDH expression was used as a loading control. Representative experiment of 4 performed. P < 0.05 was considered significant.
