Fig. 5. Receptor tyrosine kinase activation in response to growth factor stimulation. Receptor tyrosine kinase activation in response to growth factor.

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Fig. 5. Receptor tyrosine kinase activation in response to growth factor stimulation. Receptor tyrosine kinase activation in response to growth factor stimulation. (A) HDMEC, HCMEC, HBMEC and A2780 were grown to sub-confluence and treated with either VEGF-A or NRG-1 at 50 ng/ml for 10 min. Intracellular signalling responses were analysed by western blotting for phosphorylation of VEGFR-2, HER2 (EGFR-2), AKT and ERK1/2 using phospho-specific antibodies. Total protein was measured for VEGFR-2, HER2 and HER4. Level of protein phosphorylation and protein levels are quantified relative to HDMEC basal level. (B) Cells were treated with 50 ng/ml VEGF-A or 50 ng/ml NRG-1 for 72 h. Cell proliferation was assessed by ATP Cell Titer Glo ATP assay. Data is presented as % of basal for each cell type, mean±s.d. (n=3), *P≤0.05 compared to basal, one-way ANOVA, SPSS. (C) Cells were treated with 50 ng/ml NRG-1 for 72 h following pre-incubation with human IgG control (10 µg/ml) or Herceptin (10 µg/ml). Data is presented as % of basal for each cell type, mean±s.d. (n=3), *P≤0.05, one-way ANOVA, SPSS. Emma L. Wilkinson et al. Biology Open 2016;5:1362-1370 © 2016. Published by The Company of Biologists Ltd