anti p p38 mapk antibody thr180 tyr182  (Cell Signaling Technology Inc)

Journal: Journal of Evidence-based Integrative Medicine

Article Title: “Huanglianjiedu Decoction” Against Pancreatic Adenocarcinoma Proliferation of by Downregulating the PI3K/AKT/mTOR and MAPK/ERK1/2 Signaling Pathways

doi: 10.1177/2515690X241291381

Figure Lengend Snippet: PI3K/AKT/mTOR and MAPK/ERK1/2 signaling pathways may be involved in the therapeutic effect of HLJDD on PAAD. Western blot analysis of protein levels of p-mTOR, mTOR, p-AKT, AKT, p-PI3K, PI3K, p38 MAPK, p-p38 MAPK, ERK1/2, p-ERK1/2 and GAPDH in Capan-1 and Panc02 cells. All data were expressed as mean ± SD (ns p > .05; * p < .05; ** p < .01; *** p < .001; **** p < .0001).

Article Snippet: The primary antibodies used included anti-Ki-67 antibody (Cat. #ab15580, Abcam, UK), anti-p-ERK1/2 antibody (Cat. #4370, Cell Signaling Technology, CST, USA), and anti-p-p38 MAPK antibody (Thr180/Tyr182) (Cat. #4511S, CST).

Techniques: Western Blot

Journal: Journal of Evidence-based Integrative Medicine

Article Title: “Huanglianjiedu Decoction” Against Pancreatic Adenocarcinoma Proliferation of by Downregulating the PI3K/AKT/mTOR and MAPK/ERK1/2 Signaling Pathways

doi: 10.1177/2515690X241291381

Figure Lengend Snippet: Histological and immunohistochemical analysis of tumor tissues from C57BL/6 mice with subcutaneous Panc02 xenografts treated with HLJDD. Hematoxylin and Eosin (HE) staining illustrates the tumor architecture at 1× and 10× magnification across control (vehicle), 50 mg/kg, 100 mg/kg, and 200 mg/kg HLJDD-treated groups. Ki-67 immunohistochemistry staining, shown at 1× and 10× magnification, indicates cell proliferation, with reduced staining intensity in HLJDD-treated groups, suggesting decreased proliferation. Phosphorylated p38 MAPK (p-p38 MAPK) and phosphorylated ERK (p-ERK) staining at 1× and 10× magnification show decreased staining intensity with increasing HLJDD doses, indicating inhibition of the MAPK and ERK signaling pathways. Scale bars represent 2.5 mm (1×) and 200 μm (10×).

Article Snippet: The primary antibodies used included anti-Ki-67 antibody (Cat. #ab15580, Abcam, UK), anti-p-ERK1/2 antibody (Cat. #4370, Cell Signaling Technology, CST, USA), and anti-p-p38 MAPK antibody (Thr180/Tyr182) (Cat. #4511S, CST).

Techniques: Immunohistochemical staining, Staining, Control, Immunohistochemistry, Inhibition

Journal: Journal of Evidence-based Integrative Medicine

Article Title: “Huanglianjiedu Decoction” Against Pancreatic Adenocarcinoma Proliferation of by Downregulating the PI3K/AKT/mTOR and MAPK/ERK1/2 Signaling Pathways

doi: 10.1177/2515690X241291381

Figure Lengend Snippet: Rescue experiment demonstrating the effects of HLJDD on the PI3K/AKT/mTOR and MAPK signaling pathways in Capan-1 and Panc02 cells. (A) Immunoblot analysis of PI3K, p-PI3K, AKT, p-AKT, mTOR, and p-mTOR in Capan-1 and Panc02 cells treated with HLJDD (200 μg/mL) in the presence or absence of the PI3K activator 740 Y-P (30 μM). HLJDD treatment reduces the levels of phosphorylated PI3K, AKT, and mTOR, indicating inhibition of the PI3K/AKT/mTOR pathway. The addition of 740 Y-P partially reverses this inhibition, restoring phosphorylation levels. (B) Immunoblot analysis of p38 MAPK, p-p38 MAPK, ERK1/2, and p-ERK1/2 in Capan-1 and Panc02 cells treated with HLJDD (200 μg/mL) in the presence or absence of the PKC activator PMA (500 nM). HLJDD treatment reduces the levels of phosphorylated p38 MAPK and ERK1/2, indicating inhibition of the MAPK pathway. The addition of PMA partially reverses this inhibition, restoring phosphorylation levels. GAPDH serves as a loading control in all experiments.

Article Snippet: The primary antibodies used included anti-Ki-67 antibody (Cat. #ab15580, Abcam, UK), anti-p-ERK1/2 antibody (Cat. #4370, Cell Signaling Technology, CST, USA), and anti-p-p38 MAPK antibody (Thr180/Tyr182) (Cat. #4511S, CST).

Techniques: Western Blot, Inhibition, Control

Journal: Journal of Proteome Research

Article Title: Comparison of Quantitative Mass Spectrometric Methods for Drug Target Identification by Thermal Proteome Profiling

doi: 10.1021/acs.jproteome.3c00111

Figure Lengend Snippet: Losmapimod interacts with MAPK14 and reduces p38 MAPK phosphorylation. (A) Assessment of losmapimod binding to MAPK14 using surface plasmon resonance (SPR). Representative multicycle SPR sensograms for losmapimod showing a dose-dependent concentration series against immobilized MAPK14. K D , equilibrium dissociation rate constant; K on (Ms –1 ), on-rate constant or association reaction; K off (s –1 ), off-rate constant or dissociation reaction. Experiments were performed in triplicate. (B) Western blot analysis of MAPK14 and p38 MAPK phosphorylation in THP-1 cells with the vehicle control, DMSO, and 1 μM losmapimod for 1 h shows loss of p38 phosphorylation in response to losmapimod. GAPDH serves as a loading control. A representative image of three replicates is shown. Relative mobilities of reference proteins (kDa) are shown on the left of each blot.

Article Snippet: Membranes were probed with the following antibodies purchased from Cell Signaling Technology: MAPKAPK3 (#7421), MAPK14 (#9218), Thr180/Tyr182-P-p38 MAPK (#4511), and anti-rabbit IgG-HRP (#7074).

Techniques: Binding Assay, SPR Assay, Concentration Assay, Western Blot