Figure 5.

The channel-dependent function of TRPA1 is not required for its contribution to both the FGF2-mediated enhancement of the EMT signature and cell invasion. PANC-1 and MIA PaCa-2 cells were pre-treated with the selective TRPA1 pore function inhibitor (A-967079) for 1 h and then left untreated or stimulated with FGF2 for 24 h, as shown above. (A) Western blot analysis showed that the inhibition of the TRPA1 pore function did not impact PANC-1 cell response to FGF2 in terms of E-cadherin/vimentin modulation toward EMT enhancement. The densitometric analysis and the statistical evaluation were performed as reported in the Materials and Methods section; * p < 0.05; *** p < 0.001. (B) Immunofluorescence analysis showed that the effects of FGF2 on PANC-1 cells, in terms of changes in cell morphology and the increased intensity of vimentin immunostaining, were not affected by the TRPA1 inhibitor. Bar: 10 μm. Quantitative analysis of cell circularity and the statistical evaluation were performed as reported above; ** p < 0.01.