Journal: International Journal of Molecular Sciences

Article Title: The Effect of Valine on the Synthesis of α-Casein in MAC-T Cells and the Expression and Phosphorylation of Genes Related to the mTOR Signaling Pathway

doi: 10.3390/ijms26073179

Figure Lengend Snippet: Effects of different valine concentrations on the phosphorylation of downstream proteins in the mTOR signaling pathway. ( A ) Western blotting was used to detect the protein expression levels of phosphorylated mTOR (P-mTOR) and phosphorylated S6K1 (P-S6K1). ( B ) The phosphorylation level of mTOR protein (P-mTOR/mTOR) was quantified and calculated using β-actin and mTOR as internal references. ( C ) The phosphorylation level of S6K1 protein (P-S6K1/S6K1) was quantified and calculated using β-actin and S6K1 as internal references. ( D ) Western blotting was used to detect the protein expression levels of phosphorylated 4EBP1 (P-4EBP1) and phosphorylated RPS6 (P-RPS6). ( E ) The phosphorylation level of 4EBP1 protein (P-4EBP1/4EBP1) was quantified and calculated using β-actin and 4EBP1 as internal references. ( F ) The phosphorylation level of RPS6 protein (P-RPS6/RPS6) was quantified and calculated using β-actin and RPS6 as internal references. ( G ) Western blotting was used to detect the protein expression levels of phosphorylated eEF2 (P-eEF2). ( H ) The phosphorylation level of eEF2 protein (P-eEF2/eEF2) was quantified and calculated using β-actin and eEF2 as internal references. The error bars represent the SD ( n = 3). ANOVA was used to detect the differences between groups, and the LSD method was combined for uncorrected exploratory pairwise comparisons, while the Duncan method was used for conservative multiple comparison corrections. Different lowercase letters indicate significant differences ( p < 0.05), while different uppercase letters indicate significant differences ( p < 0.01).

Article Snippet: , RPS6 , biorbyt , orb585017 , 5% egg white , 1:1000.

Techniques: Phospho-proteomics, Western Blot, Expressing, Comparison

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Valine on the Synthesis of α-Casein in MAC-T Cells and the Expression and Phosphorylation of Genes Related to the mTOR Signaling Pathway

doi: 10.3390/ijms26073179

Figure Lengend Snippet: Effects of valine and rapamycin on the phosphorylation levels of mTOR downstream proteins. ( A ) The phosphorylation levels of mTOR (P-mTOR) in MAC-T cells treated with varying concentrations of rapamycin were assessed using Western blotting. ( B ) The P-mTOR level (P-mTOR/mTOR) in MAC-T cells treated with 100 nM rapamycin was quantified and calculated using β-actin and mTOR as internal references. ( C ) Western blotting was used to detect the protein expression levels of P-mTOR and P-S6K1. ( D ) The P-mTOR level (P-mTOR/mTOR) was quantified and calculated using β-actin and mTOR as internal references. ( E ) The P-S6K1 level (P-S6K1/S6K1) was quantified and calculated using β-actin and S6K1 as internal references. ( F ) Western blotting was used to detect the protein expression levels of P-4EBP1 and P-RPS6. ( G ) The P-4EBP1 level (P-4EBP1/4EBP1) was quantified and calculated using β-actin and 4EBP1 as internal references. ( H ) The P-RPS6 level (P-RPS6/RPS6) was quantified and calculated using β-actin and RPS6 as internal references. ( I ) Western blotting was used to detect the protein expression levels of P-eEF2 and α-casein. ( J ) The P-eEF2 level (P-eEF2/eEF2) was quantified and calculated using β-actin and eEF2 as internal references. ( K ) The α-casein protein expression level was quantified using β-actin as an internal reference. The error bars represent the SD ( n = 3). ANOVA was used to detect the differences between groups, and the LSD method was combined for uncorrected exploratory pairwise comparisons, while the Duncan method was used for conservative multiple comparison corrections. Different lowercase letters indicate significant differences ( p < 0.05), while different uppercase letters indicate significant differences ( p < 0.01).

Article Snippet: , RPS6 , biorbyt , orb585017 , 5% egg white , 1:1000.

Techniques: Phospho-proteomics, Western Blot, Expressing, Comparison

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Valine on the Synthesis of α-Casein in MAC-T Cells and the Expression and Phosphorylation of Genes Related to the mTOR Signaling Pathway

doi: 10.3390/ijms26073179

Figure Lengend Snippet: Primer sequences of internal reference genes and target genes.

Article Snippet: , RPS6 , biorbyt , orb585017 , 5% egg white , 1:1000.

Techniques: Sequencing

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Valine on the Synthesis of α-Casein in MAC-T Cells and the Expression and Phosphorylation of Genes Related to the mTOR Signaling Pathway

doi: 10.3390/ijms26073179

Figure Lengend Snippet: Detailed information of antibodies.

Article Snippet: , RPS6 , biorbyt , orb585017 , 5% egg white , 1:1000.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Effect of Valine on the Synthesis of α-Casein in MAC-T Cells and the Expression and Phosphorylation of Genes Related to the mTOR Signaling Pathway

doi: 10.3390/ijms26073179

Figure Lengend Snippet: Detailed information of antibodies.

Article Snippet: , RPS6 , biorbyt , orb585017 , 5% egg white , 1:1000.

Techniques:

Journal: Oncology Reports

Article Title: Evaluation of co‑inhibition of ErbB family kinases and PI3K for HPV‑negative head and neck squamous cell carcinoma

doi: 10.3892/or.2025.8871

Figure Lengend Snippet: A combination of copanlisib and afatinib effectively blocked the phosphorylation of HER2, HER3 and Akt, and enhanced caspase-3 cleavage. (A) Cal27 and (B) FaDu cells were treated with increasing concentrations of copanlisib, 0.5 µM afatinib or their combination for 24 h before lysis. The indicated proteins were detected by western blot analysis. C-, cleaved; P-, phosphorylated.

Article Snippet: The following antibodies were purchased from Cell Signaling Technology, Inc.: Phosphorylated (P)-Akt-S473 (cat. no. 4058), P-Akt-T308 (cat. no. 9275), Akt (cat. no. 2938), P-HER2-Y1248 (cat. no. 2247), HER2 (cat. no. 4290), P-HER3-Y1289 (cat. no. 2842), HER3 (cat. no. 12708), caspase-3 (cat. no. 9665), cleaved-caspase-3 (cat. no. 9664) and β-actin (cat. no. 4967).

Techniques: Phospho-proteomics, Lysis, Western Blot

Journal: Oncology Reports

Article Title: Evaluation of co‑inhibition of ErbB family kinases and PI3K for HPV‑negative head and neck squamous cell carcinoma

doi: 10.3892/or.2025.8871

Figure Lengend Snippet: A combination of copanlisib and afatinib effectively blocked the phosphorylation of HER2, HER3 and Akt, and enhanced caspase-3 cleavage. (A) Cal27 and (B) FaDu cells were treated with increasing concentrations of copanlisib, 0.5 µM afatinib or their combination for 24 h before lysis. The indicated proteins were detected by western blot analysis. C-, cleaved; P-, phosphorylated.

Article Snippet: The following antibodies were purchased from Cell Signaling Technology, Inc.: Phosphorylated (P)-Akt-S473 (cat. no. 4058), P-Akt-T308 (cat. no. 9275), Akt (cat. no. 2938), P-HER2-Y1248 (cat. no. 2247), HER2 (cat. no. 4290), P-HER3-Y1289 (cat. no. 2842), HER3 (cat. no. 12708), caspase-3 (cat. no. 9665), cleaved-caspase-3 (cat. no. 9664) and β-actin (cat. no. 4967).

Techniques: Phospho-proteomics, Lysis, Western Blot