Fig. 6

VEGF and MEK1/2 signalling is required for endothelial cell survival. E12.5 testes cultured for 24h with DMSO, 500nM of MEKi or 100, 500 or 2500nM of VEGFRi and analysed with immunofluorescence or flow cytometry. A IF in sections of cultured testes stained with DAPI (blue), CD31 (endothelial cells and germ cells; green), cleaved Caspase 3/9 (cCaspase 3/9; red) and EdU (cell proliferation; cyan). Key: G = gonad, M = mesonephros. Arrows indicate cCaspase 3/9 negative and EdU positive endothelial cells, asterisks indicate cCaspase 3/9 positive and EdU negative endothelial cells. B Flow cytometric analysis of non-Sertoli somatic cells with cell proliferation based on EdU incorporation. Scale bar represents 100 μm. Biological replicates; in A; n = 4 per treatment, in B; n = 6 testes per treatment. In B; ordinary one-way ANOVA with Tukey’s multiple comparisons. Data represents mean ± SEM. Only significance between treatments and DMSO or 500 nM MEKi are shown, *P < 0.05, ***P < 0.001, ****P < 0.0001