Figure 2.
Wt1 expression is necessary to maintain a mesenchymal phenotype in immortalised epicardial cells. (a) Heart of Wt1-GFP knockin mouse. Scale bar represents 50 μm. (b) Direct GFP expression on heart cryosection of Wt1-GFP knockin embryos shows GFP expression in epicardial cells. Scale bar represents 50 μm. (c) Phase-contrast micrograph of Wt1 conditional KO immortalised mouse epicardial cells (CoMEEC). Scale bar represents 100 μm. (d-f) GFP, ZO-1 and Wt1 expression in CoMEEC. Scale bars represent 50 μm. CoMEEC displayed a cobblestone monolayer typical of epicardial cells (c, e). (g) Western blot analysis was conducted in Cre+ tamoxifen inducible CoMEEC cultured in the presence of different concentrations of tamoxifen. (h) RT-PCR analyses of Snai1 and Cdh1 expression in Cre+ CoMEEC in presence of tamoxifen. (i) The migratory behaviour of Cre+ CoMEEC in presence of tamoxifen was analyzed in an in vitro wound model. Scale bar represents 100 μm.