Journal: Experimental Physiology

Article Title: Skeletal muscle myosin heavy chain fragmentation as a potential marker of protein degradation in response to resistance training and disuse atrophy

doi: 10.1113/EP092093

Figure Lengend Snippet: Total myofibril protein and MyHC Total poly‐ubiquitination in 1boutRE participants. (a) Data from well‐trained 1boutRE men ( n = 8) show that total myofibril protein polyubiquitination levels remain unaltered post‐exercise. (b) Additionally, the polyubiquitination signal on immunoprecipitated MyHC fragments (spanning ∼15–50 kDa) remained unaltered 3 and 6 h post‐exercise when data were normalized to the IP: MyHC signal. (c) The Dynabead‐mouse anti‐MyHC IgG2a yielded an appreciably higher signal of putative MyHC fragments compared to a negative control (Dynabead‐mouse non‐specific IgG) when incubated with ∼600 µg myofibril isolates from the same participant sample. Representative immunoblots are shown for two of eight participants, and data are presented as means and SD values with one‐way repeated measures (RM) ANOVA P ‐values. MyHC, myosin heavy chain.

Article Snippet: These antibodies included non‐concentrated clone supernatants of: (i) mouse monoclonal IgG2a MyHC, termed MyHC Total throughout (Developmental Studies Hybridoma Bank; Iowa City, IA, USA; cat. no.: A4.1025), (ii) mouse monoclonal IgG1 MyHCI (Developmental Studies Hybridoma Bank; cat. no.: A4.951), (iii) mouse monoclonal IgG1 MyHCIIa (Developmental Studies Hybridoma Bank; cat. no.: SC‐71), and (iv) mouse monoclonal IgM MyHCIIx (Developmental Studies Hybridoma Bank; cat. no.: 6H1).

Techniques: Immunoprecipitation, Negative Control, Incubation, Western Blot