Figure 3.

TRPA1 depletion impacts on the FGFR2c-mediated enhancement of the EMT phenotype in response to FGF2. PANC-1 and MIA PaCa-2 cells were transfected with TRPA1 siRNA or control (Cx) siRNA and left untreated or stimulated with FGF2 for 24 h, as shown above. (A) Western blot analysis showed that only in PANC-1 cells, the decrease in the epithelial marker E-cadherin and the increase in the mesenchymal marker vimentin induced by FGF2 stimulation are counteracted by TRPA1 gene silencing. E-cadherin and vimentin expressions did not significantly change in MIA PaCa-2 cells. Results are expressed as mean value ± SD. The densitometric analysis and the statistical evaluation were performed as reported in the Materials and Methods section; * p < 0.05. (B) Immunofluorescence analysis showed that the effects of FGF2 stimulation in terms of changes in cell morphology (detachment from each other and acquisition of a spindle shape) and increases in intensity of vimentin immunostaining, visible only in PANC-1 cells, appeared reversed by TRPA1 depletion. Bar: 10 μm. Quantitative analysis of cell circularity and the statistical evaluation were performed as reported in the Materials and Methods section; ** p < 0.01.