Xing-Csrp3-protein measurments

To elucidate the mechanism by which CSRP3 modulates pig satellite cell differentiation, quantitative proteomics analysis of CSRP3-knockdown versus control pig satellite cells at day 2 of differentiation was performed. Proteins from pig satellite cells were extracted using RIPA buffer supplemented with PMSF and a protease inhibitor cocktail. Cell proteins were solubilized in 7M urea and 2M thiourea, and protein concentrations were quantified using the Bradford Protein Assay Kit. Proteins were digested following the FASP method. The resulting peptides were analyzed using a Q-Exactive high-resolution mass spectrometer coupled with a Nano-Acquity nano HPLC system. Data processing for peak picking and alignment was performed using Pyogenesis QI for Proteomics software (build 2.0, Nonlinear Dynamics, Newcastle, UK).