Journal: Molecular Medicine Reports

Article Title: RUVBL1 and CRAF promote periodontal ligament stem cell osteogenic differentiation via the MEK/ERK signaling cascade

doi: 10.3892/mmr.2026.13881

Figure Lengend Snippet: Overexpression of CRAF activates the RAF/MEK/ERK signaling pathway. Reverse transcription-quantitative PCR was used to analyze the mRNA expression of (A) ARAF, (B) BRAF, (C) CRAF, (D) MEK1, (E) MEK2, (F) ERK1and (G) ERK2. Western blotting was used to analyze the protein levels of (H) ARAF, (I) BRAF, (J) CRAF, (K) MEK1/2, (L) p-MEK1/2, (M) p-MEK1/2 to MEK1/2, (N) ERK1/2, (O) p-ERK1/2 and (P) p-ERK1/2 to ERK1/2. *P<0.05, **P<0.01 and ***P<0.001. p-, phosphorylated; OE, overexpression; NC, negative control; sh, short hairpin; ns, not significant; ARAF, A-Raf proto-oncogene serine/threonine-protein kinase; BRAF, B-Raf proto-oncogene serine/threonine-protein kinase; CRAF, C-Raf proto-oncogene serine/threonine-protein kinase.

Article Snippet: The antibodies were as follows: GAPDH (1:1,000; cat. no. P30008M; Abmart Pharmaceutical Technology Co., Ltd.), A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) (cat. no. bs-2251R), CRAF (cat. no. bs-23170R), MEK1/2 (cat. no. bs-1041R), phosphorylated (p)-MEK1/2 (cat. no. bs-3270R), ERK1/2 (cat. no. bsm-33232M), p-ERK1/2 (cat. no. bs-3016R; all BIOSS), RUVBL1 (cat. no. 74775; Cell Signaling Technology, Inc.), ERK1/2 (all 1:500; cat. no. bsm-33232M; BIOSS), B-Raf proto-oncogene serine/threonine-protein kinase (BRAF) (1:2,000; cat. no. ab33899; Abcam) and HRP-conjugated universal secondary antibody [cat. nos.

Techniques: Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Negative Control

Journal: Molecular Medicine Reports

Article Title: RUVBL1 and CRAF promote periodontal ligament stem cell osteogenic differentiation via the MEK/ERK signaling cascade

doi: 10.3892/mmr.2026.13881

Figure Lengend Snippet: Overexpression of RUVBL1 activates the MEK/ERK signaling pathway. Reverse transcription-quantitative PCR was used to analyze the mRNA expression of (A) ARAF, (B) BRAF, (C) CRAF, (D) MEK1, (E) MEK2, (F) ERK1 and (G) ERK2. Western blotting was used to analyze protein levels of (H) ARAF, (I) BRAF, (J) CRAF, (K) MEK1/2, (L) p-MEK1/2, (M) p-MEK1/2 to MEK1/2, (N) ERK1/2, (O) p-ERK1/2 and (P) p-ERK1/2 to ERK1/2. *P<0.05, **P<0.01 and ***P<0.001. RUVBL1, RuvB-like AAA ATPase-1; p-, phosphorylated; OE, overexpression; NC, negative control; sh, short hairpin; ns, not significant; ARAF, A-Raf proto-oncogene serine/threonine-protein kinase; BRAF, B-Raf proto-oncogene serine/threonine-protein kinase; CRAF, C-Raf proto-oncogene serine/threonine-protein kinase.

Article Snippet: The antibodies were as follows: GAPDH (1:1,000; cat. no. P30008M; Abmart Pharmaceutical Technology Co., Ltd.), A-Raf proto-oncogene serine/threonine-protein kinase (ARAF) (cat. no. bs-2251R), CRAF (cat. no. bs-23170R), MEK1/2 (cat. no. bs-1041R), phosphorylated (p)-MEK1/2 (cat. no. bs-3270R), ERK1/2 (cat. no. bsm-33232M), p-ERK1/2 (cat. no. bs-3016R; all BIOSS), RUVBL1 (cat. no. 74775; Cell Signaling Technology, Inc.), ERK1/2 (all 1:500; cat. no. bsm-33232M; BIOSS), B-Raf proto-oncogene serine/threonine-protein kinase (BRAF) (1:2,000; cat. no. ab33899; Abcam) and HRP-conjugated universal secondary antibody [cat. nos.

Techniques: Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Negative Control

Journal: Animal Nutrition

Article Title: Modulating starch digestion kinetics via feed processing: Implications for growth and metabolism in weaned pigs

doi: 10.1016/j.aninu.2025.08.011

Figure Lengend Snippet: Effect of corn processing on relative mRNA level for mTOR , 4E-BP1 , p70S6K and protein abundances of mTOR, phosphorylated mTOR (p-mTOR), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), phosphorylated 4E-BP1 (p-4E-BP1), p70 ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6K (p-p70S6K) in the longissimus thoracis of piglets. Results are presented as means ± standard error of the mean (SEM), n = 4. Data columns with different letters were significantly different ( P ≤ 0.05).

Article Snippet: The membranes were blocked at room temperature, followed by incubation at 4 °C overnight with the following primary antibodies: mammalian target of rapamycin (mTOR, catalog No. 2983, Cell Signaling Technology, Danvers, MA, USA), phosphorylated mTOR (p-mTOR, catalog No. 5536, Cell Signaling Technology, Danvers, MA, USA), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1, catalog No. 9644, Cell Signaling Technology, Danvers, MA, USA), phosphorylated 4E-BP1 (p-4E-BP1, catalog No. 2855, Cell Signaling Technology, Danvers, MA, USA), p70 ribosomal protein S6 kinase (p70S6K, catalog No. 2708, Cell Signaling Technology, Danvers, MA, USA), phosphorylated p70S6K (p-p70S6K, catalog No. 9234, Cell Signaling Technology, Danvers, MA, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, catalog No. 5174, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Binding Assay

Journal: Poultry Science

Article Title: PKA and PKC signaling pathways mediate vitamin D₃-regulated intestinal phosphorus absorption in broiler chickens

doi: 10.1016/j.psj.2026.106762

Figure Lengend Snippet: Effects of dietary vitamin D 3 supplementation on jejunal phosphorylation levels of PKA, PKC, PI3K, p38MAPK, and ERK in broiler chickens (19 d) (Experiment 1). The control and vitamin D 3 diets contained 0 and 1000 IU/kg vitamin D 3 , respectively. (a) p-PKA/t-PKA; (b) p-PKC/t-PKC; (c) p-PI3K/t-PI3K; (d) p-p38MAPK/t-p38MAPK; (e) p-ERK/t-ERK. Protein abundance values are means ± SD of four replicates per treatment (1 broiler per replicate) (n = 4). Values with different letters differ between treatments ( P < 0.05).

Article Snippet: PVDF membranes were incubated with primary antibodies against NaPi-IIb (1:1000, A9460; ABclonal, Wuhan, China), phosphorylated PKA (p-PKA; 1:1000, 5661S), phosphorylated PI3K (p-PI3K; 1:1000, 4228), phosphorylated ERK (p-ERK; 1:1000, 9101S), phosphorylated p38MAPK (p-p38MAPK; 1:1000, 9216S), total PKA (t-PKA; 1:1000, 5842S) (Cell Signaling Technology, Boston, USA), total PI3K (t-PI3K; 1:10000, 60225-1-Ig), total ERK (t-ERK; 1:8000, 11257-1-AP-50), total p38MAPK(t-p38MAPK; 1:1000, 14064-1-AP), total PKC (t-PKC; 1:1000, 21991-1-AP), phosphorylated PKC (p-PKC; 1:5000, 29123-1-AP), and GAPDH (1:10000, 60004-1-Ig) (Proteintech, Wuhan, China).

Techniques: Phospho-proteomics, Control, Quantitative Proteomics

Journal: Poultry Science

Article Title: PKA and PKC signaling pathways mediate vitamin D₃-regulated intestinal phosphorus absorption in broiler chickens

doi: 10.1016/j.psj.2026.106762

Figure Lengend Snippet: Effects of the PKA inhibitor H-89 on jejunal phosphorylation levels of PKC, PI3K, p38MAPK, and ERK in broiler chickens (10–15 d) (Experiment 2). (a) p-PKC/t-PKC; (b) p-PI3K/t-PI3K; (c) p-p38MAPK/t-p38MAPK; (d) p-ERK/t-ERK. Protein abundance values are means ± SD of four replicates per treatment (1 broiler per replicate) (n = 4). Values with different letters differ between treatments ( P < 0.05).

Article Snippet: PVDF membranes were incubated with primary antibodies against NaPi-IIb (1:1000, A9460; ABclonal, Wuhan, China), phosphorylated PKA (p-PKA; 1:1000, 5661S), phosphorylated PI3K (p-PI3K; 1:1000, 4228), phosphorylated ERK (p-ERK; 1:1000, 9101S), phosphorylated p38MAPK (p-p38MAPK; 1:1000, 9216S), total PKA (t-PKA; 1:1000, 5842S) (Cell Signaling Technology, Boston, USA), total PI3K (t-PI3K; 1:10000, 60225-1-Ig), total ERK (t-ERK; 1:8000, 11257-1-AP-50), total p38MAPK(t-p38MAPK; 1:1000, 14064-1-AP), total PKC (t-PKC; 1:1000, 21991-1-AP), phosphorylated PKC (p-PKC; 1:5000, 29123-1-AP), and GAPDH (1:10000, 60004-1-Ig) (Proteintech, Wuhan, China).

Techniques: Phospho-proteomics, Quantitative Proteomics

Journal: Poultry Science

Article Title: PKA and PKC signaling pathways mediate vitamin D₃-regulated intestinal phosphorus absorption in broiler chickens

doi: 10.1016/j.psj.2026.106762

Figure Lengend Snippet: Effects of the PKC inhibitor staurosporine on duodenal phosphorylation levels of PKA, PI3K, p38MAPK, and ERK in broiler chickens (13 d) (Experiment 3). (a) p-PKA/t-PKA; (b) p-PI3K/t-PI3K; (c) p-p38MAPK/t-p38MAPK; (d) p-ERK/t-ERK. Protein abundance values are means ± SD of four replicates per treatment (1 broiler per replicate) (n = 4). Values with different letters differ between treatments ( P < 0.05).

Article Snippet: PVDF membranes were incubated with primary antibodies against NaPi-IIb (1:1000, A9460; ABclonal, Wuhan, China), phosphorylated PKA (p-PKA; 1:1000, 5661S), phosphorylated PI3K (p-PI3K; 1:1000, 4228), phosphorylated ERK (p-ERK; 1:1000, 9101S), phosphorylated p38MAPK (p-p38MAPK; 1:1000, 9216S), total PKA (t-PKA; 1:1000, 5842S) (Cell Signaling Technology, Boston, USA), total PI3K (t-PI3K; 1:10000, 60225-1-Ig), total ERK (t-ERK; 1:8000, 11257-1-AP-50), total p38MAPK(t-p38MAPK; 1:1000, 14064-1-AP), total PKC (t-PKC; 1:1000, 21991-1-AP), phosphorylated PKC (p-PKC; 1:5000, 29123-1-AP), and GAPDH (1:10000, 60004-1-Ig) (Proteintech, Wuhan, China).

Techniques: Phospho-proteomics, Quantitative Proteomics

Journal: Animal Nutrition

Article Title: Modulating starch digestion kinetics via feed processing: Implications for growth and metabolism in weaned pigs

doi: 10.1016/j.aninu.2025.08.011

Figure Lengend Snippet: Effect of corn processing on relative mRNA level for mTOR , 4E-BP1 , p70S6K and protein abundances of mTOR, phosphorylated mTOR (p-mTOR), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), phosphorylated 4E-BP1 (p-4E-BP1), p70 ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6K (p-p70S6K) in the longissimus thoracis of piglets. Results are presented as means ± standard error of the mean (SEM), n = 4. Data columns with different letters were significantly different ( P ≤ 0.05).

Article Snippet: The membranes were blocked at room temperature, followed by incubation at 4 °C overnight with the following primary antibodies: mammalian target of rapamycin (mTOR, catalog No. 2983, Cell Signaling Technology, Danvers, MA, USA), phosphorylated mTOR (p-mTOR, catalog No. 5536, Cell Signaling Technology, Danvers, MA, USA), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1, catalog No. 9644, Cell Signaling Technology, Danvers, MA, USA), phosphorylated 4E-BP1 (p-4E-BP1, catalog No. 2855, Cell Signaling Technology, Danvers, MA, USA), p70 ribosomal protein S6 kinase (p70S6K, catalog No. 2708, Cell Signaling Technology, Danvers, MA, USA), phosphorylated p70S6K (p-p70S6K, catalog No. 9234, Cell Signaling Technology, Danvers, MA, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, catalog No. 5174, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Binding Assay

Journal: Poultry Science

Article Title: Heterologous expression, immunogenic evaluation, and subunit vaccine potential of the σC protein from the Xinjiang avian reovirus (ARV) strain xj-1.1

doi: 10.1016/j.psj.2026.106779

Figure Lengend Snippet: Standard curve for phosphorylation level determination of recombinant σC proteins.

Article Snippet: Phosphorylation levels were quantified using the formula provided by the Phosphorylation Assay Kit (purchased from Sangon Biotech (Shanghai) Co., Ltd., catalog number: C500061): N (Protein phosphorylation level%) = (X × 2.24% × A)/(B × 31) Where: N: Molar percentage of phosphorus per mole of the target phosphorylated protein; A: Molecular weight of the target protein (g/mol); B: Total protein concentration of the sample (μg/mL); X: Phosphorylated protein concentration of the sample (μg/mL); Note: The phosphorylation level of the phosphorylated standard protein used in this assay was 2.24%.

Techniques: Phospho-proteomics, Recombinant

Journal: Animal Nutrition

Article Title: Modulating starch digestion kinetics via feed processing: Implications for growth and metabolism in weaned pigs

doi: 10.1016/j.aninu.2025.08.011

Figure Lengend Snippet: Effect of corn processing on relative mRNA level for mTOR , 4E-BP1 , p70S6K and protein abundances of mTOR, phosphorylated mTOR (p-mTOR), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), phosphorylated 4E-BP1 (p-4E-BP1), p70 ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6K (p-p70S6K) in the longissimus thoracis of piglets. Results are presented as means ± standard error of the mean (SEM), n = 4. Data columns with different letters were significantly different ( P ≤ 0.05).

Article Snippet: The membranes were blocked at room temperature, followed by incubation at 4 °C overnight with the following primary antibodies: mammalian target of rapamycin (mTOR, catalog No. 2983, Cell Signaling Technology, Danvers, MA, USA), phosphorylated mTOR (p-mTOR, catalog No. 5536, Cell Signaling Technology, Danvers, MA, USA), eukaryotic initiation factor 4E-binding protein 1 (4E-BP1, catalog No. 9644, Cell Signaling Technology, Danvers, MA, USA), phosphorylated 4E-BP1 (p-4E-BP1, catalog No. 2855, Cell Signaling Technology, Danvers, MA, USA), p70 ribosomal protein S6 kinase (p70S6K, catalog No. 2708, Cell Signaling Technology, Danvers, MA, USA), phosphorylated p70S6K (p-p70S6K, catalog No. 9234, Cell Signaling Technology, Danvers, MA, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, catalog No. 5174, Cell Signaling Technology, Danvers, MA, USA).

Techniques: Binding Assay