Fig. 5

Targeting of FGFR1 signaling by combination treatment with AZD4547 and GDC0941. SH5YSY and SKNBE2 cells were transiently transfected with pCMV6-FGFR1^wt, pCMV6-FGFR1^N546K and pCMV6 empty vector. A, B Total protein extracts were analyzed by western blotting to evaluate the levels of phosphorylated and total FGFR1, STAT3, ERK and AKT. The β-Actin protein levels were used as loading control. C The ability of cells treated with single AZD4547 0.1 µM and with combination of AZD4547 0.1 µM and GDC0941 1 µM to invade and migrate through a matrigel-coated membrane support was evaluated. The number of invading FGFR1^wt or FGFR1^N546K overexpressing cells are shown in percentage respect to untreated cells (100% vehicle). D The ability of cells to form neuropheres after treatment with AZD4547 0.1 µM alone and in combination with GDC0941 1 µM was evaluated. The colony number of FGFR1^wt or FGFR1^N546K overexpressing cells are shown in percentage respect to untreated cells (100% vehicle). Vehicle = DMSO; p = p-value