Figure 4

(A) Embryos were injected at 0-cell stage with either mrc1a:wt-hKRAS, mrc1a:hKRAS p.Gly12Asp, mrc1a:hKRAS p.Gly13Asp and tol2 transposase. Larvae at 7 dpf under light microscopy (2.5× magnification), GFP (2.5× magnification), mCherry (2.5× magnification), and confocal microscopy (20× magnification). Larvae injected with mrc1a:hKRAS p.Gly12Asp or p.Gly13Asp under light microscopy have edema around the heart and intestine (arrows). Mrc1a:wt-hKRAS have essentially normal vasculature of larvae injected with under confocal microscopy. Vasculature of larvae injected with mrc1a:hKRASp.Gly12Asp or mrc1a:hKRASp.Gly13Asp under confocal microscopy showing fusion of the thoracic duct with the cardinal vein (brackets). (B) In vivo zebrafish larvae therapeutic screening. Embryos were treated at 48 hpf and screened for edema at 5.5 dpf. (C) Fraction of larvae with edema by KRAS variant, drug, and concentration. Each dot represents a single experiment. *P < 0.05 by unpaired, 1-tailed Student?s t tests, after correction for multiple testing with the Benjamini and Hochberg FDR method. The mechanism of action for each drug can be seen in Supplemental Table 1. Due to figure legend space limitations, the number of zebrafish larvae for each experiment are in Supplemental Table 2.