Label-Free LC-MS/MS Proteomics Analyses Reveal Proteomic Changes AccompanyingMSTNKO in C2C12 Cells

Abstract

Analysis of the proteome of myostatin (MSTN) knockout (KO) mouse C2C12 cells has proven valuable to studies investigating the molecular mechanisms by which MSTN regulates skeletal muscle development. To identify new protein/pathway alterations and candidate biomarkers for skeletal muscle development, we compared proteomic profiles of MSTN KO C2C12 cells (KO) with corresponding wild-type cells (NC) using a label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique. A total of 2637 proteins were identified and quantified in KO cells. Among these proteins, 77 proteins were significantly differentially expressed, 38 upregulated, and 39 downregulated, in MSTN KO C2C12 cells. These significantly altered proteins are involved in metabolic processes, developmental processes, immune system processes, and the regulation of other biological processes. Enrichment analysis was utilized to link these alterations to biological pathways, which are predominantly related to oxidative phosphorylation, protein digestion and absorption, mitochondrion localisation, antigen processing and presentation, the MAPK signaling pathway, the PPAR signaling pathway, the PI3K-Akt signaling pathway, and the JAK-STAT signaling pathway. Upregulation of several proteins, including epoxide hydrolase, tropomyosin 1, Cyb5a, HTRA1, Cox6a1, CD109, Synap29, and Ugt1a6, likely enhanced skeletal muscle development, the immune system, and energy metabolism. Collectively, our results present a comprehensive proteomics analysis of MSTN KO C2C12 myoblast cells; we hypothesize that MSTN KO could activate p38MAPK signaling pathway by CDC42, and we further deciphered the function of MSTN in the regulation of skeletal muscle development, immune processes, and mitochondrial energy metabolism.

IRE1α regulates skeletal muscle regeneration through myostatin mRNA decay

“…We then determined whether Myostatin is required for IRE1α-mediated effect on myoblast differentiation using Mstn-KO C2C12 cells in which Mstn was abrogated by the CRISPR/Cas9-based strategy (45). Immunostaining of MyHC in differentiated myotubes showed slightly increased fusion index and myotube diameter in Mstn-KO cells relative to the wildtype (WT) control cells (Figure 5A-C…”

Proteomic Profiling Reveals Differentially Expressed Proteins Associated with Amylose Accumulation During Rice Grain Filling

“…Proteomics pro ling has been approved as a powerful molecular strategy that has been widely implemented in dissecting the molecular basis of various biological processes in living organisms including plants. However, the experimental system and procedure of the employed proteomics pro ling approach greatly affect the powerful and e ciency of proteomic pro ling in dissecting the molecular mechanisms of a biological process [18][19][20][21][22][23][24][25][26]. In rice, seed development is a complex biological process that is greatly affect grain yield and quality and is governed by complex regulatory networks comprising numerous transcription factors [27].…”

Comparative Quantitative Proteomic Analysis Reveals Differentially Expressed Proteins Involved in Grain-Filling Rate of Rice at the Early Ripening Stage

“…Analysis of proteomics pro les has been demonstrated as a powerful and e cient molecular tool in exploring molecular mechanisms underlying various biological processes in living organisms. However, the e ciency of proteomic analysis in identi cation of proteins underlying a given biological process greatly depends on the applied proteomics analysis procedure and the experimental system [28][29][30][31][32][33]. Although proteomic analyses have been widely employed to dissect the molecular basis of variable biological processes in plants, in particular stress tolerance, [29,32,[34][35][36][37][38][39]), to the best of our knowledge this is the rst comprehensive study conducted to dissect proteins involved in grain lling rate of rice.…”