Fig. 4

ppp1r10 inhibition rescues aberrantly activated Erk signaling in dA/+. (A) Schematics of ppp1r105’UTR generated by MMEJ sgRNA. The underlined sequence indicates sgRNA sequence, red indicates deleted nucleotides. (B) Evaluation of ppp1r10 and ppp2caa gene transcripts expression by quantitative RT-PCR in hearts from heterozygous ppp1r10+/− and ppp2ca+/− adult zebrafish at 3 months old. n = 4. t-test was used to compare 2 groups. Data are presented as mean ± SEM, *p < 0.05, **p < 0.01. (C) Representative western blots of protein Erk, pErk, Rsk, pRsk and Gapdh from 3 months old zebrafish hearts and quantification of Erk, pErk, Rsk, and pRsk. One-way ANOVA was used to compare multiple groups for each mutation. Data are presented as mean ± SEM, three biological replicates, **p < 0.01. WT, wild type, ns, non-significant. (D) Representative images and quantification of pErk in cryosectioned heart tissues of WT, dA/+, ppp2caa+/−; dA/+ and ppp1r10+/−; dA/+ zebrafish mutants co-immunostained using anti-pErk antibody and anti-α-actinin antibody. α-actinin, green. pErk, red. DAPI, blue. The open arrowheads indicate pErk. pErk, phosphorylated Erk1/2. Scale bars: 20 μm. One-way ANOVA was used to compare multiple groups for each mutation. Data are presented as mean ± SEM, three biological replicates, **p < 0.01. WT, wild type, ns, non-significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)