Fig. S2

Expression of kat2a and kat2b in is reduced in mutants; genotyping strategy for kat2a and kat2b mutants, heterozygotes and WTs. Lateral views of in situ hybridization for (A) kat2a in kat2a-/- and (B) kat2b in kat2b-/- zebrafish at 36 hpf. (C) Genotyping PCR to distinguish between kat2a wildtype, heterozygotes (het, +/-) and mutants (-/-). PCR was performed with primers designed to amplify the TALEN target region in exon 1 of kat2a. PCR products were digested with SacI, an enzyme whose target sequence that lies within the TALEN site, and whose sequence is altered in the TALEN mutants. Band patterns of digested products indicate their genotype as labeled. (D) Genotyping PCR to validate mutagenesis of kat2b by CRISPR/Cas9. Left panel shows PCR amplification of kat2b exon 6 CRISPR target region using primers targeted to that region. PCR followed by T7 Endonuclease I yields the respective band patterns as labeled. Right panel shows PCR amplification of kat2b exon 6 CRISPR target region using a dCaps forward primer designed against the mutant sequence. Digestion of PCR product by AciI yields the respective band patterns as labeled. Scale bars represent 250 μm.