Fig 2. Transgenic mice had human mutant MYOC expressed in skeletal muscle and human MYOC is detected by Western blot as a doublet due to partial N-glycosylation.

(A) Western blots for human MYOC protein in adult transgenic mouse gastrocnemius muscle lysates. Human MYOC protein was detected in the transgenic mouse skeletal muscle and in the heart using R&D Systems anti-MYOC antibody (1:1000). Loading was 40μg tissue lysate per well of a 10% SDS-PAGE gel. Total mouse number for this representative western blot is N = 6 different animals. (B) NTM5 cells were transiently-transfected with a plasmid with cDNA for human MYOC-FLAG or a plasmid with cDNA for mouse Myoc-FLAG. 20μg cell lysates were treated with PNGase to remove N-glycosylation followed by Western blot analysis using anti-FLAG (1:1000). (C) Weights of gastrocnemius muscles and hearts harvested from equal numbers of male and female wt and CMV-MYOC-Y437H transgenic mice did not differ between groups. N per group was ≥ 6. +/- SD is indicated and t-test were p>0.1. Abbreviations–wild-type, wt; transgenic, Tg.