Fig. 2. Kermit 2 loss-of-function phenotype. (A) A kermit 2-directed antisense morpholino blocks translation of kermit 2-GFP mRNA (5'UTR-kermit 2-GFP, which contains the morpholino target sequence in the 5'UTR), but not kermit 2-GFP mRNA lacking the 5'UTR. Kermit 2-GFP mRNA (1 ng) with or without the 5'UTR was injected into one-cell embryos together with kermit 2 morpholino (K2M; 20 ng). Embryos were harvested at stage 10 and analyzed by western blot with GFP antibodies. ß-Tubulin was used as a loading control. (B) Unilateral injection of kermit 2 morpholino (40 ng) into one dorsal blastomere at the four-cell stage completely blocked eye development in 60% of embryos (lower left panel, n=38) and reduced eye formation in an additional 32% (not shown). Co-expression of kermit2-GFP mRNA, which lacked the morpholino target sequence, restored eye formation in 72% of kermit2-depleted embryos (lower right panel, n=43). kermit2-GFP mRNA alone did not cause apparent embryonic defects (upper right panel).