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Fig. 2. Immunodetection of RyR1. (A) ED13 MA and PM muscle fibers with and without innervation were immunostained with an anti-RyR1 antibody and an FITC conjugated secondary antibody. Control for nonspecific immunostaining by the secondary antibody is included. (B) Protein extracts from ED13 PM and MA muscles in vivo as well as PM and MA muscle fibers cultured in the absence (-SC) and presence (+SC) of innervation were electrophoresed and blotted. RyR1 was detected using an anti-RyR1 antibody. {alpha}-Actin was used as a loading control for normalization of RyR1 abundance and was detected using an anti-sarcomeric {alpha}-actin antibody. (C) Western blots of RyR1 in extracts from ED13 PM and MA muscles in vivo as well as PM and MA muscle fibers cultured in the absence (-SC) and presence (+SC) of innervation were quantitated. PM muscle fibers contained significantly more RyR1 relative to MA muscle fibers (*P<0.03; **P<0.01). Bars represent means±s.e.m.; n=5.





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