phosphorylated p38mapk  (Cell Signaling Technology Inc)

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: Frontiers in Physiology

Article Title: Direct simulation of hypertensive stress on endothelial cells: a streamlined model of in-vitro-hypertension

doi: 10.3389/fphys.2025.1724932

Figure Lengend Snippet: Intra-cellular signaling pathways in unstimulated or LPS-stimulated HUVEC seeded in LB1, cultured in static or dynamic conditions. NFkB (A) and p38MAPK (B) phosphorylation after stimulation with LPS (1 μg/mL). Endothelial cells seeded in LB1 for 24 h were incubated with LPS or medium alone both in static and under flow for further 24 h, as experimental control. Results are expressed as the ratio of phosphorylated to non-phosphorylated form, normalized to the respective control and evaluated using Image J software. Western Blotting images are representative of a single experiment. Histograms represent mean of three independent experiments. pNFkB: phosphorylated NFkB; p38MAPK: phosphorylated p38MAPK.

Article Snippet: The membranes were blocked for 2 h at room temperature (RT) in PBS/0.05% Tween 20 (PT) (Cell Signaling Technology) containing 5% non-fat dry milk (Santa Cruz Biotechnology, Dallas, TX, United States) and incubated overnight at 4 °C with anti-human NFκB (1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #8242) or anti-human phosphorylated NFκB (pNFκB, 1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #3039) or anti-human p38MAPK (1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #9212) or anti-human phosphorylated p38MAPK (pp38MAPK, 1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #9211) After three washes, the membranes were incubated for 1 h at RT in PT/5% non-fat dry milk plus HRP-conjugated secondary antibodies (1:5000, Cell Signaling Technology, catalog number #7074) and developed using ECL (Westar Supernova, Cyanagen, Bologna, IT, catalog number XLS3,0100).

Techniques: Protein-Protein interactions, Cell Culture, Phospho-proteomics, Incubation, Control, Software, Western Blot

Journal: Frontiers in Physiology

Article Title: Direct simulation of hypertensive stress on endothelial cells: a streamlined model of in-vitro-hypertension

doi: 10.3389/fphys.2025.1724932

Figure Lengend Snippet: Intra-cellular signaling pathways in unstimulated or Angiotensin II-stimulated HUVEC seeded in LB1, cultured in static or dynamic conditions. NFkB (A) and p38MAPK (B) phosphorylation after stimulation with ANG II (1,000 nM). Endothelial cells seeded in LB1 for 24 h were incubated with Angiotensin II or medium alone both in static and under flow for further 24 h. Results are expressed as the ratio of phosphorylated to non-phosphorylated form, normalized to the respective control and evaluated using Image J software. Western Blotting images are representative of a single experiment. Histograms represent mean of three independent experiments. pNFkB: phosphorylated NFkB; p38MAPK: phosphorylated p38MAPK.

Article Snippet: The membranes were blocked for 2 h at room temperature (RT) in PBS/0.05% Tween 20 (PT) (Cell Signaling Technology) containing 5% non-fat dry milk (Santa Cruz Biotechnology, Dallas, TX, United States) and incubated overnight at 4 °C with anti-human NFκB (1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #8242) or anti-human phosphorylated NFκB (pNFκB, 1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #3039) or anti-human p38MAPK (1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #9212) or anti-human phosphorylated p38MAPK (pp38MAPK, 1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #9211) After three washes, the membranes were incubated for 1 h at RT in PT/5% non-fat dry milk plus HRP-conjugated secondary antibodies (1:5000, Cell Signaling Technology, catalog number #7074) and developed using ECL (Westar Supernova, Cyanagen, Bologna, IT, catalog number XLS3,0100).

Techniques: Protein-Protein interactions, Cell Culture, Phospho-proteomics, Incubation, Control, Software, Western Blot

Journal: Frontiers in Physiology

Article Title: Direct simulation of hypertensive stress on endothelial cells: a streamlined model of in-vitro-hypertension

doi: 10.3389/fphys.2025.1724932

Figure Lengend Snippet: Intra-cellular signaling pathways in HUVEC seeded in LB1, cultured in static or dynamic conditions, unstimulated or stimulated with Angiotensin II and Live-PA, individually or in combination. NFkB (A) and p38MAPK (B) phosphorylation after stimulation with ANG II (1,000 nM) alone or in combination with Live-Pa. Endothelial cells seeded in LB1 for 24 h were stimulated with ANG II under flow for 24 h. Live-Pa was applied for further 2 h in the presence of medium or ANG II. Results are expressed as the ratio of phosphorylated to non-phosphorylated form, normalized to the dynamic control and evaluated using Image J software. Western Blotting images are representative of a single experiment. Histograms represent mean of three independent experiments. pNFkB: phosphorylated NFkB; p38MAPK: phosphorylated p38MAPK.

Article Snippet: The membranes were blocked for 2 h at room temperature (RT) in PBS/0.05% Tween 20 (PT) (Cell Signaling Technology) containing 5% non-fat dry milk (Santa Cruz Biotechnology, Dallas, TX, United States) and incubated overnight at 4 °C with anti-human NFκB (1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #8242) or anti-human phosphorylated NFκB (pNFκB, 1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #3039) or anti-human p38MAPK (1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #9212) or anti-human phosphorylated p38MAPK (pp38MAPK, 1:1,000 in PT plus 5% BSA, Cell Signaling Technology, catalog number #9211) After three washes, the membranes were incubated for 1 h at RT in PT/5% non-fat dry milk plus HRP-conjugated secondary antibodies (1:5000, Cell Signaling Technology, catalog number #7074) and developed using ECL (Westar Supernova, Cyanagen, Bologna, IT, catalog number XLS3,0100).

Techniques: Protein-Protein interactions, Cell Culture, Phospho-proteomics, Control, Software, Western Blot

Journal: Scientific Reports

Article Title: The pro-tumor activity of INTS7 on lung adenocarcinoma via inhibiting immune infiltration and activating p38MAPK pathway

doi: 10.1038/s41598-024-77093-3

Figure Lengend Snippet: Potential mechanisms involved in the regulatory activity of INTS7. (A) Immunohistochemical staining and statistical analysis of INTS7 in clinical tumor tissues ( n = 6, Low = Group with low INTS7 expression, High = Group with high INTS7 expression, scale bar: 50 μm). (B) Immunohistochemical staining and statistical analysis of cleaved caspase-3 (c-caspase-3) in clinical tumor tissues ( n = 6, Low = Group with low INTS7 expression, High = Group with high INTS7 expression, scale bar: 50 μm). (C) H&E staining of xenograft tumor tissues (scale bar: 50 μm). (D) Immunohistochemical staining and statistical analysis of INTS7 in xenograft tumor tissues ( n = 5, scale bar: 50 μm). (E) Immunohistochemical staining and statistical analysis of cleaved caspase-3 (c-caspase-3) in xenograft tumor tissues ( n = 5, scale bar: 50 μm). (F) Expression of INTS7 by western blot analysis ( n = 3). (G) Expression of cleaved caspase-3 (c-caspase-3), p53, phosphorylated p38MAPK (p-p38MAPK) and MMP2 by western blot analysis ( n = 3, respectively). The grouping of blots were cropped from different parts of the same gel. Original images of blots are presented in Supplementary Figs. 2–6). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Membranes were blocked with 5% bovine serum albumin (BSA) and incubated overnight at 4 ℃ with the following primary antibodies: anti-INTS7 (1:1000, Proteintech, USA), anti-cleaved caspase-3 (1:1000, Proteintech, USA), anti-p53 (1:500, Proteintech, USA), anti-p38MAPK (1:1000, Servicebio, China), anti-phosphorylated p38MAPK (1:1000, Servicebio, China), anti-MMP2 (1:1000, Servicebio, China). β-actin (1:5000, Proteintech, USA) was used as loading control.

Techniques: Activity Assay, Immunohistochemical staining, Staining, Expressing, Western Blot