Journal: Scientific Reports

Article Title: Acute resistance exercise and training reduce desmin phosphorylation at serine 31 in human skeletal muscle, making the protein less prone to cleavage

doi: 10.1038/s41598-024-79385-0

Figure Lengend Snippet: Desmin phosphorylation in untrained condition at resting state (R1) in healthy young human skeletal muscle. Qualitative assessment. Pooled muscle samples ( n = 3) were fractionated in a detergent (Triton X-100) -soluble supernatant (S) and insoluble pellet (P). Samples were applied as duplicates (I, II) and membranes were incubated with antibodies raised against desmin phosphorylated at serine 31 (pS31) ( B ), 60 (pS60) ( C ), threonine 17 (pT17) ( D ), 76/77 (pT76/77) ( E ) as well as total desmin ( A , F ; two different antibodies; one monoclonal [mono] and one polyclonal [poly]). Ponceau S was used as loading standard. Vertical arrowheads indicate bands or fraction positive for respective antibody. Horizontal arrowheads indicate the analyzed bands. It is to note that the p Des S60 antibody recognized a single band which however appeared slightly above the other bands (see text). For p Des S76/77 , only bands at predicted height were considered.

Article Snippet: Primary: Desmin (D93F5), total (rabbit monoclonal IgG AB; #5332S; 1:1500; Cell Signaling Technology, Danvers, MA, USA); Desmin, total (rabbit polyclonal IgG AB; #PA5-16705; 1:3000; Thermo Fisher Scientific, Waltham, MA, USA); Desmin, phospho threonine 17 (rabbit polyclonal igG AB; #bs-5301R; 1:750; Bioss Antibodies, Woburn, MA, USA); Desmin, phospho serine 31 (rat monoclonal igG2b; #D375-3; 1: 2250; MBL International Corporation, Woburn, MA, USA); Desmin, phospho serine 60 (rabbit polyoclonal igG AB; #PA5-38837; 1:500; Thermo Fisher Scientific, Waltham, MA, USA); secondary: anti-rabbit IgG, HRP-linked (#7074S; 1:7500; Cell Signaling Technology, Danvers, MA, USA); anti-rat igG2b, HRP-linked (#PA1-84710; 1:7500; Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Phospho-proteomics, Incubation