Fig. 3 S2

Characterization of Tg(-0.8flt1:tdTomato) expression during development, and validation of Tg(hsp70l:sflt1, cryaa:cerulean) and Tg(hsp70l:sflt4, cryaa:cerulean) lines.

(A–A”, B–B”, and C–C”) Confocal images of Tg(-0.8flt1:tdTomato);Tg(kdrl:EGFP) trunks at 34 (A–A”), 56 (B–B”), and 80 (C–C”) hpf. Tg(kdrl:EGFP) expression labels all the ISVs, whereas Tg(-0.8flt1:tdTomato) expression labels aISVs strongly and vISVs weakly at 80 hpf. Scale bar, 100 µm. (D–G) 32 hpf Tg(kdrl:EGFP);Tg(hsp70l:sflt1, cryaa:cerulean) (D and E), Tg(kdrl:EGFP) (F), and Tg(kdrl:EGFP);Tg(hsp70l:sflt4, cryaa:cerulean) (G) trunk vasculature visualized by Tg(kdrl:EGFP) expression. Animals were given no heat shock (D) or single heat shock at 20 hpf (E–G). Heat shock treated Tg(kdrl:EGFP);Tg(hsp70l:sflt1, cryaa:cerulean) animals show a loss of ISVs (E). Scale bar, 100 µm. (H–K) 60 hpf Tg(fli1a:EGFP);Tg(hsp70l:sflt4, cryaa:cerulean) (H and K), Tg(fli1a:EGFP) (J), and Tg(fli1a:EGFP);Tg(hsp70l:sflt1, cryaa:cerulean) (I) trunk vasculature visualized by Tg(fli1a:EGFP) expression. Animals were given no heat shock (H) or multiple heat shocks at 32, 40, and 50 hpf (I–K). Red arrows point to the parachordal chains (H–J), and heat shock treated Tg(fli1a:EGFP);Tg(hsp70l:sflt4, cryaa:cerulean) animals show a loss of the parachordal chains (K). Scale bar, 100 µm. (L–O) 7 dpf TgBAC(etv2:EGFP);Tg(hsp70l:sflt4, cryaa:cerulean) (L and O), TgBAC(etv2:EGFP) (N), and TgBAC(etv2:EGFP);Tg(hsp70l:sflt1, cryaa:cerulean) (M) trunk vasculature visualized by TgBAC(etv2:EGFP) expression. Animals were given no heat shock (L) or multiple heat shocks starting at 32 hpf and every 8–12 hr after that (M–O). Red arrows point to the thoracic duct (L–N), and heat shock treated TgBAC(etv2:EGFP);Tg(hsp70l:sflt4, cryaa:cerulean) animals show a loss of the thoracic duct (O). Scale bar, 100 µm.