Fig. 6 | Nature Communications

Fig. 6

From: High mitogenic stimulation arrests angiogenesis

Fig. 6

Inhibition of ERK phosphorylation decreases EC sprouting and proliferation. a, b Confocal micrographs of retinas treated with IgG (control) or anti-Dll4 antibody in the presence or absence of MEK/ERK inhibitor (SL327) for 24 h. P-ERK immunostaining is strongly reduced in SL327-treated retinas, leading to decreased sprouting (n = 3 mice/6 retinas per group). c Confocal micrographs of retinas treated with IgG (control) or anti-Dll4 antibody in the presence or absence of SL327 for 24 h and immunostained for ERG (red) and EdU (blue) to check effect of ERK signalling on EC proliferation. Dual ERG+/EdU+ cells are pseudocolored in green. d, e Inhibition of ERK phosphorylation with SL327 for 24 h, significantly reduces the frequency of EdU+/ERG+ ECs at the AF and the anti-Dll4 induced proliferation in the mature vascular area (n = 3 mice/6 retinas per group). f, g Confocal micrographs of retinas treated with IgG (control) or anti-Dll4 antibody in the presence or absence of SL327 for 24 h and immunostained for ERG (red) and p21 (green). Quantifications show that SL327 treatment for 24 h reduced the frequency of p21+ ECs, with a particularly strong inibitory effect on the p21 upregulation induced by anti-Dll4 (n = 3 mice/6 retinas per group). Scale bars in all panels, 200 μm. Error bars represent StDev. *p < 0,05, ***p < 0.0005 and NS not significant. One-way ANOVA with Tukey’s post hoc test. Source data are provided as a Source Data file

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