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Fig. 7 PP1β paralogs are required for proper zebrafish body axis elongation.
(A–I) Lateral views of representative 48 hpf zebrafish embryos injected with (A) uninjected control, (B) 2.5 ng ppp1cba MO, (C) 2.5 ng ppp1cbb MO, (D) a mixture of 0.75 ng ppp1cba MO and 0.75 ng of ppp1cbb MO (2MO), (E) 200 pg of ppp1cba mRNA, (F) 200 pg of ppp1cbb mRNA (G) a partially rescued embryo injected with 100 pg ppp1cba mRNA and 2.5 ng of ppp1cbb MO (H) a partially rescued embryo injected with 100 pg ppp1cbb mRNA and 2.5 ng of ppp1cbb MO (I) a partially rescued embryo injected with 100 pg ppp1cbb mRNA and 0.75 ng of ppp1cbb and 0.75 ppp1cba MO (J) Quantification of the truncated body axis phenotype in morphant and mRNA injected embryos. Each injection was performed multiple times with 50 embryos used to calculate body axis length and reported as % of uninjected clutch mates. Error bars are standard error and a black * indicates a statistically significant difference from control and a # indicates a statistically significant rescue compared to the corresponding morpholino injected embryos. Statistical significance was calculated using a one-factor ANOVA with Tukey post hoc analysis and is defined as p < 0.05. (K) A western blot showing PP1β levels in zebrafish embryo lysates from control and embryos injected with two doses of morpholino cocktail (1.5 ng and 3.0 ng of total MO), individual ppp1cba or ppp1cbb (2.5 ng of either) morpholinos or a rescued embryo injected with 0.75 ng of each morpholino (1.5 ng total) and 100 pg of ppp1cbb mRNA. Tubulin was used as a loading control.