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Fig. S3 Gβ proteins are expressed during zebrafish embryonic development. (A) Western blot analysis showing the expression of Gβ proteins at 80% epiboly in embryos injected with 4MO (a combination of four MOs that target gnb1a, gnb1b, gnb4a and gnb4b, 4 ng each) was decreased, when compared with that in control uninjected embryos. α-Tubulin was used as a loading control. An anti-pan Gβ antibody (1:5,000, Santa Cruz Biotechnology) and an anti-tubulin antibody (1:10,000, Sigma) were used. (B-E′) Confocal images of wild-type embryos and 4MO-injected embryos immunostained with a pan-Gβ antibody (1:200). Maternally deposited Gβ proteins were present in embryos at the 64-cell and sphere stages (B,C). Expression of Gβ proteins at shield stage and 80% epiboly was not significantly reduced in 4MO-injected embryos (D′,E′). Scale bars: 200 μm in B; 10 μm in C-E2.