Fig. 6

Effects of VEGF stimulation in Marimastat-induced amoeboid conditions on signaling molecules. A Confocal microscopy for pERM (red fluorescence) and RhoA (green fluorescence) under mesenchymal (CTRL) and amoeboid (Marimastat) conditions, in the absence and in the presence of VEGF (25 ng/ml) ± Bevacizumab (5 µg/ml). Morphological features of the mesenchymal (elongated) to amoeboid (roundish) transition of ECFCs are evident in the enlarged insets. Nuclear staining: DAPI (blue). Colocalization between pERM and RhoA: MERGE. Magnification ×40 for reference pictures and ×100 for enlarged insets. The shown pictures are representative of 30 different pictures for each experimental condition. B–G Western blotting results show the effects of VEGF, in mesenchymal and amoeboid conditions in the absence and in the presence of VEGF ± Bevacizumab, on the intracellular signaling molecules RhoA and Rac1, the phosphorylation of ERK and MLC2, WAVE2, uPAR and Ephrin-A1. Numbers on the right refer to molecular weights expressed in kDa. Histograms report band densitometry. Results are the mean of 5 different experiments performed in duplicate and are shown as mean value ± SD. *p < 0.05; **p < 0.001; ***p < 0.0001 significantly different from control