Fig. 3.

Icotinib enhanced the inhibition effect of pemetrexed in migration and invasion in a sequence-dependent manner. a, b Representative images and quantification of the results of wound healing assay in PC-9 (a) and A549 cells (b). The magnification of the microscope is 40×. Data are presented as mean ± SD of triplicate experiments. c, d Representative images and quantification of the results of transwell invasion assay in PC-9 (c) and in A549 cells (d). The magnification of the microscope is 100×. e The EMT related protein molecules were analyzed by western blot in PC-9 and A549 cells treated with pemetrexed with or without icotinib. f Cytoskeleton was analyzed by immunofluorescence assay for phalloidin (red) with DAPI counterstaining (blue) in PC-9 cells treated with pemetrexed with or without icotinib (400×). Data are presented as mean ± SD of triplicate experiments, *p < 0.05