Journal: Animal Nutrition
Article Title: Myotonic dystrophy-related CDC42-binding kinase alpha (MRCKα) mediates methionine- and leucine-stimulated β-casein synthesis in bovine mammary epithelial cells via targeting mTOR
doi: 10.1016/j.aninu.2025.01.003
Figure Lengend Snippet: Silencing of MRCKα inhibits the stimulatory effects of Met and Leu on mRNA abundance of genes related to milk protein synthesis in bovine mammary epithelial cells. Cells were transfected with scrambled siRNA (control, SC) or MRCKα siRNA (SI) and stimulated simultaneously with control (B, basal medium), additional Met (M, Met = 0.6 mM) or Leu (L, Leu = 0.6 mM). After 24 h, cells were collected and isolated for RT-qPCR analysis. Values are presented as mean ± SEM for three independent experiments. P AA , P -value of amino acid addition (AA); P SI , P -value of silencing of MRCKα (SI); P AA×SI , P -value of AA × SI interaction. CSN2 = β-casein; CSN1S1 = αS1-casein; CSN3 = κ-casein; LALBA = lactalbumin α; MRCKα = myotonic dystrophy-related CDC42-binding kinase alpha; mTOR = mechanistic target of rapamycin; JAK2 = Janus kinase 2; STAT5 = signal transducers and activators of transcription 5.
Article Snippet: Mouse monoclonal anti-human PKB antibody, mouse monoclonal anti-human phosphorylated PKB (Thr 308 ) antibody, rabbit polyclonal anti-mouse Janus kinase 2 (JAK2) antibody and rabbit polyclonal anti-mouse phosphorylated JAK2 (Tyr 1007/1008 ) antibody were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).
Techniques: Transfection, Control, Isolation, Quantitative RT-PCR, Binding Assay