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. 2020 Jul 17;11:3606. doi: 10.1038/s41467-020-17384-1

Fig. 6. Nebivolol inhibits endothelial cells metabolism and proliferation.

Fig. 6

a Representative western blots of two different samples and quantification of four different samples (histograms) of the expression of VEGF and VEGFR2 (*p = 0.01) in tumors of control (CRL) and nebivolol-treated (NEB) colon xenograft mice. Tubulin is shown as loading control. b–h HUVEC cells were treated (NEB, red dots and bars) or not (CRL, closed dots and bars) during 3 or 48 h with 10 µM nebivolol. b Immunofluorescence microscopy images of VEGFR2 (VEGFR2, green) and DAPI (blue)-stained HUVEC treated as indicated. Scale bar, 50 µm. Histograms represent the relative expression of VEGFR2-positive cells relative to cell nuclei from nine different slices. c Immunofluorescence microscopy images of VEGFR2 (VEGFR2, green) and DAPI (blue)-stained HUVEC treated as indicated. Scale bar, 50 µm. Histograms represent the number of cells from nine different slices. *p = 0.03. d The figure shows the proliferation after 48 h of CellTrace Red Far incorporation. The blue curve shows time 0. The plot shows the quantification of cellular proliferation of four different samples. **p = 3.1E−10. See Supplementary Fig. 7c for gating strategy. e Representative western blot of the expression of β1-adrenergic receptor (ADRB1) in cancer and in human umbilical vein endothelial cells (HUVEC). Tubulin is shown as loading control. Western blots were repeated three times with similar results. f Glycolytic flux measured by the initial rates of lactate production of six different biological samples are shown. **p = 0.005. g Plot shows the percentage of cells in G0/G1, S and G2/M phases of the cell cycle from three different samples. See Supplementary Fig. 7d for gating strategy. h Representative western blot of the expression of ERK and its phosphorylated form (pERK). Western blots were repeated two times with similar results. Bars indicate the mean of the indicated samples ± SEM. *p < 0.05 and **p < 0.01 when compared to CRL by two-sided Student’s t test. See also Supplementary Fig. 6. Source data are provided as a Source Data file.