Fig. S3

Fig. S3

Validation of pxna/b ATG-MO Efficacy by Western Blotting and Rescue Experiments. (A) Targeting diagram for pxna/b ATG-MO. Sequence similarity between exon 1 (E1) and exon 2 (E2) of pxna and pxnb transcripts allows for translation blocking of both genes using one MO. Alignment shows MO target sequence compared with pxna and pxnb transcripts. Yellow highlights nucleotides shared between all sequences while blue highlights those shared between two sequences. (B) Western blotting with Paxillin antibody revealed robust knockdown of endogenous Pxn proteins in embryos injected with pxna/b ATG-MO and expression of exogenous GFP-Pxna fusion protein in embryos injected with pxna/b ATG-MO + GFP-Pxna mRNA at 3 dpf. (C) Pxn protein knockdown resulted in myotome defects which included wide MTJ chevron angles and MTJ breaches (arrows), these phenotypes were partially rescued by GFP-Pxna expression that localized to MTJs. Scale bar =50 µm.