Figure 3.4

Vegfr-signalling is not required for rapid Erk activation following vessel wounding. (A?B??) Lateral spinning disc confocal images of ISV endothelial cells (ECs) in 28 hours post-fertilisation (hpf) EC-EKC embryos treated for an hour with either 0.5% dimethyl sulfoxide (DMSO) (A?A??), with active EC Erk-signalling, or 500 nM AV951 (B?B??), with inactive EC Erk-signalling. Images (A) and (B) show the fli1aep:EKC expression, while images (A?) and (B?) show both the fli1aep:EKC and the fli1a:H2B-mCherry expression. Images (A??) and (B??) show the nuclear fli1aep:EKC intensity. (C) Quantification of nucleus/cytoplasm EKC intensity in ISV tip ECs of 28 hpf embryos treated with either 0.5% DMSO (0.849, 65 ECs, n = 14 embryos) or 500 nM AV951 (1.423, 53 ECs, n = 12 embryos). (D?Z?) Vegfr-signalling inhibitors do not block rapid Erk-signalling activation in ablated and adjacent ISVs following vessel wounding. Lateral spinning disc confocal images of ISV ECs in 4 days post-fertilisation (dpf) EC-EKC larvae treated with either 0.5% DMSO (D?I?), 15 ?M SL327 (J?M?), 4 ?M SU5416 (O?R?), 10 ?M SU5416 (S?V?), or 500 nM AV951 (W?Z?). Images (D-E?) show non-ablated control ISV ECs. Images (F-G?), (J-K?), (O-P?), (S-T?), and (W-X?) show ablated ISV ECs. Images (H-I?), (L-M?), (Q-R?), (U-V?), and (Y-Z?) show adjacent ISV ECs. Images (F, H, J, L, O, Q, S, U, W, Y) were taken pre-ablation and images (G, I, K, M, P, R, T, V, X, Z) were taken 15 min post-ablation (mpa). Images (D-Z) show the fli1aep:EKC expression and images (D?-Z?) show the nuclear fli1aep:EKC intensity. White dotted lines show the wounded sites of each larva. ISV: intersegmental vessel. Statistical test: Mann-Whitney test was conducted for graph (C). Error bars represent standard deviation. Scale bars: 25 ?m for image (A), 15 ?m for image (D).