Figure 2.

TRPA1 depletion interfered with FGFR2c oncogenic signaling. PANC-1 and MIA PaCa-2 cells were transiently transfected with TRPA1-specific siRNA or unrelated siRNA (Cx siRNA) as the negative control and then left untreated or stimulated with FGF2 to induce FGFR2c activation and signaling. The efficiency of gene silencing was assessed by real-time RT-PCR (A) and Western blot (B); * p < 0.05; ** p < 0.01. Biochemical analysis showed that TRPA1 depletion did not affect FGFR2c expression at either the mRNA (C) or protein (D) level. (E) Western blot analysis showed that only in PANC-1 cells, TRPA1 silencing repressed the FGF2-mediated phosphorylation of the FGFR2c signaling platform FRS2 and that of PKCε and ERK1/2, as well as that of MTOR and its substrate S6K. 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; *** p < 0.001.