Chemical inhibition of ERK blocks ISV sprouting. (A-C) Transmitted light images of embryos treated with the indicated compound and concentration from 10?hpf to 24?hpf. Arrows in B indicate necrosis. (D) Western blot analysis of pERK, total ERK and actin in embryos treated from 24 to 25?hpf as indicated. (E-H) Transmitted light images (E,F) and confocal images (G,H) of Tg(cdh5:gal4ff)mu101;(uas:nTom)um152;(uas:egfp)nkuasgfpa1 embryos at 32?hpf. Embryos were treated with 0.1% DMSO (E,G) or 15?µM SL327 (F,H) from 20 to 32?hpf. DA and PCV are denoted by brackets in G,H and shorter ISVs are indicated by arrows in H. (I-K) Quantification of ISV length (I), cell number per ISV (J) and BrdU-positive cells per ISV (K) following treatment with the indicated compound. *P<0.05, ***P<0.001; ns, not statistically significant; numbers of ISVs and embryos counted across three experiments are indicated on the x-axis. Error bars represent s.d.
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