Fig. 4

Relevance of the FPR1 to migration of melanoma cells. a Representative images of human melanoma A375 and M14 cells immune-stained with anti-FPR1 polyclonal antibody and visualized by a fluorescence inverted microscope. Nuclei were stained blue with DAPI. Scale bar: 5 μm. Original magnification: 1000 x. b Whole cell lysates (40 μg/sample) from A375 and M14 cells were resolved on a 10% SDS-PAGE followed by Western blotting with 1 μg/mL anti-FPR1 polyclonal antibody or 0.2 μg/mL anti-GAPDH polyclonal antibody as loading controls. c A375 and M14 melanoma cells exposed to 10 nM FITC-fMLF for 30 min at 37 °C and then visualized using a Zeiss 510 Meta LSM microscope in 2D (left) or 3D (right) projections. Scale bar: 5 μm. Original magnification: 630×. d-e A375 (red) and M14 (green) melanoma cells were exposed to diluents (None), or desensitized (dotted boxes) with 100 nM fMLF (d) or 100 nM SRSRY (e) for 1 h at 37 °C and then allowed to migrate in Boyden chambers for 4 h at 37 °C toward serum-free medium (CTRL), 10% FBS or 10 nM SRSRY. The basal value assessed in the absence of chemoattractant (CTRL) was taken as 100% and all values were reported relative to that. Data are the means ± SD of three independent experiments, performed in triplicate. Statistical significance with **p < 0.001 and *** p < 0.0001