Figure 4.

MS-275 promotes ZO-1 plasma membrane localization, α-SMA downregulation and the reversal to an epithelial-like morphology independently of treatment with TGFβ1. Effect of MS-275 treatment on ZO-1 and α-SMA expression and localization. (A) MCs were treated with MS-275 (250 nM), MC2500 (250 nM) or DMSO for 72 hours. Moreover, cells were left untreated or stimulated with TGFβ1 for additional 24 hours. Only TGFβ1-treated cells are shown. Cells were fixed, permeabilized and stained with a polyclonal antibody against ZO-1. Images were acquired by confocal microscopy. Cell nuclei are shown in blue (Hoechst 33342). Confocal images are shown from one representative experiment of three performed. (B) MCs were treated as in (A). Cells were stained with a monoclonal antibody against α-SMA. Confocal images are shown from one representative experiment of three performed. (C) Effect of MS-275 on MMT induced by exposure to PD fluid. Epithelial-like MCs treated for four days with stay safe balance 4.25% and then treated with the same PD fluid in the presence of MS-275 for three more days. Alternatively, MCs were treated with stay safe balance 4.25% for seven days or left untreated for four days and then treated with DMSO for the last three days. Cells were fixed, permeabilized and stained with phalloidin and with a polyclonal antibody against ZO-1. Images were acquired by confocal microscopy. Cell nuclei are shown in blue (Hoechst 33342). Confocal images are shown from one representative experiment of three performed. Overlay images are shown. Single fluorescences are shown in Suppl. Fig. 6. P < 0.05 was considered significant.