p stat3 s727  (Cell Signaling Technology Inc)

Journal: Frontiers in Immunology

Article Title: PD0325901 alleviates thrombin-inhibited osteogenic differentiation through an IL-1β-activated feedback loop between MEK-Erk1/2 and NF-κB signal pathways: insights from bioinformatics and experimental verification

doi: 10.3389/fimmu.2026.1730337

Figure Lengend Snippet: The mutual regulatory effects of MEK-Erk1/2 and Stat3 following treatment with PD03 and C188-9, and the role of Stat3 pathways in osteogenic differentiation. (A, B) The phosphorylation levels of Erk1/2, Stat3 (Y705), and Stat3 (S727) in osteoblasts were examined by western blot following treatment with thrombin and PD03 at different time points, and the relative protein levels of p-Erk1/2 to Erk1/2, p-Stat3(Y705) to Stat3, and p-Stat3(S727) to Stat3 were measured using Image J software. (C, D) The protein levels of p-Erk1/2, p-Stat3 (Y705), and p-Stat3 (S727) in osteoblasts were analyzed by western blot after being treated with thrombin and C188–9 at different time points, and the relative protein levels of p-Erk1/2 to Erk1/2, p-Stat3(Y705) to Stat3, and p-Stat3(S727) to Stat3 were evaluated using Image J software. (E) Osteoblasts were stained for ALP and Fluo-4 AM after being induced with the osteogenic differentiation medium for 7 days. (F) An enzymatic assay quantified ALP activities in cell lysates. (G) Western blot analysis was used to examine the expression levels of Col1α1, Runx2 and OCN. (H) The relative protein levels of Col1α1/β-actin, Runx2/β-actin and OCN/β-actin were measured using ImageJ software. (I) The expression levels of Col1α1, Runx2, Osterix, OPG, and OCN were assessed by qPCR following thrombin and C188–9 treatment for 7 days. Data are presented as mean ± SD (n = 3). P-values were determined by one-way ANOVA (multi-group comparisons) (*p < 0.05; **p < 0.01; ***p < 0.001; ns, P >0.05). Scale bar: 100 μm.

Article Snippet: The following antibodies were used in this study: β-actin (AF5001, Beyotime, China), Col1α1 (ab270993, Abcam, UK), Runx2 (AF2593, Beyotime, China), OCN (AF6297, Beyotime, China), Igf1 (AF7179, Beyotime, China), Wnt5a (29793-1-AP, Proteintech, USA), Tgfb3 (AF8142, Beyotime, China), Spp1 (A5427, Bimake, USA), COX-2 (F0327, Bimake, USA), Matrix Metalloproteinase-9 (MMP-9, AF5234, Beyotime, China), Minichromosome Maintenance Complex Component 2 (MCM2, A5172, Bimake, USA), Proliferating Cell Nuclear Antigen (PCNA, SC-25280, Santa, USA), PAR-1 (AF6837, Beyotime, China), IL-1RA (AF7218, Beyotime, China), p65 (8242S, CST, USA), p-p65 (3033T, CST, USA), Erk1/2 (4695T, CST, USA), p-Erk1/2 (4370S, CST, USA), p-Stat3 (Y705) (9145S, CST, USA), p-Stat3 (S727) (9134S, CST, USA), and Stat3 (4904T, CST, USA).

Techniques: Phospho-proteomics, Western Blot, Software, Staining, Enzymatic Assay, Expressing

Journal: Journal of Inflammation Research

Article Title: Anti-Inflammatory Regulatory Role of Signal Transducer and Activator of Transcription 3 Phosphorylation in Regulating Hypersensitivity Responses to Echinococcus granulosus Hydatid Cyst Fluid

doi: 10.2147/JIR.S509286

Figure Lengend Snippet: The inhibitory effect of different concentrations of inhibitors on STAT3 phosphorylation.

Article Snippet: The STAT3 inhibitors Stattic and JSI-124 were procured from Shanghai Haoyuan Chemexpress Co., Ltd. Antibodies for STAT3, phosphorylated p-STAT3 (S727), p-STAT3 (Y705), and ACTIN, as well as secondary antibodies, were purchased from Wuhan ABclonal Biotechnology Co., Ltd. High-resolution rapid electrophoresis liquid and ice-free rapid transfer liquid were obtained from Wuhan Servicebio Technology Co., Ltd. Immunoglobulin E (IgE) was purchased from Sigma-Aldrich, USA.

Techniques: Phospho-proteomics