Fig. S8

PI3K inhibitor wortmannin or MEK inhibitor U0126 could induce hematopoietic and angiogenic defects similar to the defects induced by K-ras knock-down.

(A) Either wortmannin or U0126 treatment were able to cause the hematopoietic defects. These defects include empty heart, with no or few red blood cells inside heart (indicated by arrows in ii and iii, compared to wild type in i), reduced or lack of normal circulation and reduced number of circulating red blood cells (indicated by arrows in v and vi, compared to wild type in iv), and accumulation of blood cells in some sites away from the circulation (indicated by arrows in vii and viii). All embryos were observed at 4 dpf (days-post fertilization), lateral view, anterior to the left and dorsal to the top.

(B) o-Dianisidine staining for wortmannin or U0126 treated embryos, showing loss or reduction of hemoglobin positive cells overall, especially inside heart and in yolk sac (indicated by empty arrows and block arrows respectively in ii, iii, v and vi, compared to wild type embryos in i and iv). Except for grouped embryos, all other embryos are lateral view, anterior to the left and dorsal to the top. Embryos were observed at 6 dpf.

(C) Either wortmannin or U0126 treatment were able to cause angiogenic defects. Inhibitor treatment for fli1-GFP embryos resulted in disorganized blood vessels, including the missing segmental vessels and/or bearing ectopic vessel sprouts (indicated by arrows in i and ii), similar to the defects caused by K-ras knock-down. Embryos were observed at 4 dpf, lateral view, anterior to the left and dorsal to the top.