Fig. 3

P311 induced a myofibroblast-like phenotype in primary mouse EpSCs. a Immunofluorescence for α-SMA (upper), vimentin (middle), and E-cadherin (lower) in EpSCs at 72 hours after P311 transfection (left) and control vector transfection (right). α-SMA, vimentin, and E-cadherin were labeled using AF594 (red), and nuclei were stained with DAPI (blue). Representative images from four independent experiments are shown. Scale bar, 25 μm. b Immunofluorescence for F-actin in EpSCs. TGFβ1 served as the positive control. F-actin was labeled by Rhodamine (red), and nuclei were stained with DAPI (blue). Details for the observed cell morphologies are shown highlighted in the interior box and illustrated in the lower panel. Scale bar, 20 μm. c The mRNA level of α-SMA was determined in P311-expressing cells and control vector-expressing cells using real-time PCR (n = 4). d Western blot analysis for α-SMA, vimentin, β1-integerin, and E-cadherin in P311-transfected and control vector-transfected EpSCs. Representative blots from four independent experiments are shown in the top panel. Bottom panel shows the quantification of protein levels, which was obtained using densitometry (n = 4). Data are represented as mean ± SEM, Student’s t test, *P < 0.05 and **P < 0.01 in the P311-transfected group vs the control vector-transfected group (Color figure online)