Fig. 3

scCM promotes in vitro angiogenesis mediated by MAPK and VEGFR2 activation. scCM induces a rapid and consistent increase in ERK1/2 phosphorylation, whereas it does not affect AKT or VEGFR2 activation (a). ERK1/2 activation remains until 4 h after treatment, while VEGFR2 phosphorylation is upregulated after 24 h of treatment (b). Pre-incubation with UO126 (MEK/ERK inhibitor—1 μmol/L), VEGFR2 kinase inhibitor VII (10 μmol/L) and FGFR tyrosine kinase inhibitor (1 μmol/L) reduced the scCM-induced proliferation (c) while VEGFR2 and FGFR inhibition reduced the scCM-induced migration (d). Western blot detection of ERK1/2 and VEGFR2 phosphorylation on cells treated with either vehicle (DMSO), MEK/ERK1/2, or VEGFR2, or FGFR inhibitors (e). Proteins classified as involved in angiogenesis [8], VEGF-A [4], and FGF [15] were grouped, then converted in gene identifiers and an interaction network was prepared using Cytoscape software (version 3.8.2, released 2020.10.24). Such pathways share in common the protein MAPK1 (ERK2) (f). Data represent median (interquartile range) (n = 3–7), *P < 0.05 vs scEBM; $P < 0.05 vs scCM vehicle