Fig. 3.

Over-expression of PITX2 affects markers for EMT and invasion in ovarian cancer cells. a-d The expression level of SNAI1, SNAI2, MMP9, CLD4, CLD7, DSM and VIM genes were quantified by Q-PCR assay after ectopic over-expression of PITXA/B/C into SKOV-3 (a and c) and OAW-42 (b and d) cells. The expression of CDH1 was assessed in SKOV-3 cells (Fig A-ii) upon transient transfection of PITX2 isoforms. The comparative expression of respective genes is shown as relative ‘fold’ change (mean ± S.E.M). e-f Western blot analysis of the proteins with respective antibodies was performed with the lysates of PITX2-tranfected SKOV-3 (e) and OAW-42 (f) cells. Here, GAPDH was used as internal loading control. g Bright field microscopy images exhibit the epithelial phenotype of the control and mesenchymal phenotype of PITX2A-transfected IOSE (G-i) and SKOV3 (G-ii) cells cells. h Phalloidin staining of IOSE cells (i) and imunofluorescence imaging of E-cadherin was performed for the SKOV-3 cells (ii). i-j Q-PCR assay was performed to check the expression of CLDN1, CLDN7, CDH2 and VIM in PITX2-overexpressed IOSE cells. *represents p < 0.05