bovine b-casein c6905  (Millipore)

Journal: Metabolites

Article Title: Methionine Promotes Milk Protein Synthesis via the PI3K-mTOR Signaling Pathway in Human Mammary Epithelial Cells.

doi: 10.3390/metabo13111149

Figure Lengend Snippet: Figure 2. Effect of methionine on the expression of genes related to milk protein synthesis. MCF-10A cells were treated with different concentrations of methionine (0, 0.3, 0.6, 0.9, and 1.2 mM) for 24 h: (A–C) Effect of different concentrations of methionine on the mRNA expression levels of CSN2 (A), CSN3 (B), CSN1S1 (C); (D) CSN2 protein levels were measured by Western blotting analysis; (E) relative folds of β-casein (CSN2)/β-actin from the Western blotting were quantified by grayscale scan; (F) immunofluorescence observation of the plasma membrane localization of CSN2 in cells subjected to a 24-h treatment with 0.6 mM methionine. CSN2 (green), Hoechst (blue). The scale bar is 100 µm. Data were the means ± SEM from three independent experiments. All data were analyzed using ordinary one-way ANOVA multiple comparisons and Tukey’s multiple comparison test. Values with different superscripted lowercase letters indicate a significant difference (p < 0.05). The Met in the graphs all represent methionine.

Article Snippet: The cells were covered with primary antibody beta casein (CSN2) (Proteintech, Wuhan, China) overnight at 4 ◦C.

Techniques: Expressing, Western Blot, Clinical Proteomics, Membrane, Comparison

Journal: Metabolites

Article Title: Methionine Promotes Milk Protein Synthesis via the PI3K-mTOR Signaling Pathway in Human Mammary Epithelial Cells.

doi: 10.3390/metabo13111149

Figure Lengend Snippet: Figure 5. Effect of methionine on the expression of genes related to milk protein synthesis. MCF-10A cells were treated with different concentrations of methionine (0, 0.3, 0.6, 0.9, and 1.2 mM) for 24 h: (A) mTOR and PI3K protein levels were measured by Western blotting analysis; (B,C) relative folds of mTOR and PI3K protein levels (protein/β-actin) from the Western blotting were quantified by grayscale scan, effect of PI3K inhibition on the role of milk protein synthesis, MCF-10A cells were treated with XL-147 (477 nM) to inhibit PI3K; (D–F) effect of indicated treatment on the mRNA expression levels of CSN2, CSN3, and CSN1S1; (G–I) CSN2, mTOR, and p-mTOR protein levels were measured by Western blotting analysis; (J–L) relative folds of CSN2, mTOR, and p-mTOR protein levels (protein/β-actin) from the Western blotting were quantified by grayscale scan. Data were the means ± SEM from three independent experiments. All data were analyzed using ordinary one-way ANOVA multiple comparisons and Tukey’s multiple comparison test. Values with different superscripted lowercase letters indicate a significant difference (p < 0.05). The Met in the graphs all represents methionine.

Article Snippet: The cells were covered with primary antibody beta casein (CSN2) (Proteintech, Wuhan, China) overnight at 4 ◦C.

Techniques: Expressing, Western Blot, Inhibition, Comparison