p p38 mapk polyclonal antibody  (Proteintech)

Journal: Redox Biology

Article Title: Stromal cell-derived itaconate promotes endometriosis via macrophage NRF2 and lysosomal pH modulation

doi: 10.1016/j.redox.2026.104101

Figure Lengend Snippet: Itaconate Regulates Lysosomal Function, Calcium Signaling, and p38 MAPK Pathway in Macrophages (A) Lysosomal acidification in ascites-derived macrophages from non-endometriosis (NC) and endometriosis (EM) patients assessed by LysoSensor fluorescence intensity (n = 6/group). (B) Lysosomal acidification in PBMC-derived macrophages after LPS,IL-4 and 4-OI treatment (n = 3/group). (C) Lysosomal pH value of BMDMs after LPS and 4-OI treatment (n = 3/group). (D, E) Effect of chloroquine (CQ) on LysoSensor intensity (D) and proportion of iNOS + BMDMs (E) after LPS and 4-OI treatment (n = 3/group). (F) mRNA levels of Il1b , Il6 , Nos2 , and Tnf in BMDMs under indicated treatments (n = 3/group). (G) Intracellular Ca 2+ dynamics in BMDMs after treatments, measured by Fluo-4. (H) Intracellular Ca 2+ dynamics in ascites-derived macrophages from NC and EM patients. (I) Quantification of intracellular calcium in peritoneal macrophages from NC and EM patients (n = 3/group). (J) Quantification of intracellular calcium in BMDMs (n = 5/group). (K) mRNA expression of lysosomal calcium channel genes ( MCOLN1 , MCOLN2 , TPC1 , TPC2 ) in ascites-derived macrophages from NC and EM patients (n = 3/group). (L) lysosomal calcium channel genes mRNA in PBMC-derived macrophages co-cultured with normal ESCs (Nor-ESC), eutopic ESCs (Eu-ESC), or ectopic ESCs (Ec-ESC) from patients (n = 5/group). (M − N) Fluo-4-based Ca 2+ dynamics in BMDMs treated with LPS, 4-OI, ML-SA1 (M), or CQ (N). (O) Flow cytometry analysis and quantification of iNOS + BMDMs after indicated treatments (n = 3/group). (P) mRNA levels of pro-inflammatory genes in BMDMs under different treatments (n = 3/group). (Q) Western blot and quantification of p-p38 and total p38 in BMDMs with LPS ± 4-OI (n = 3/group). (R–S) Western blot and quantification of iNOS, p-p38, and total p38 in BMDMs treated with LPS, 4-OI, and CQ (R), or ML-SA1 (S) (n = 3/group). (Data are presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.) EM, endometriosis; NC, non-EM control; LPS, lipopolysaccharide; 4-OI, 4-octyl itaconate; CQ, chloroquine; ML-SA1, MCOLN channel agonist; PBMC, peripheral blood mononuclear cell; ESC, endometrial stromal cell.

Article Snippet: After blocking in 5% milk in TBST, membranes were incubated overnight at 4 °C with primary antibodies: Beta Actin Monoclonal antibody(1:20000, 66009-1-Ig; Proteintech), Beta Tubulin Recombinant antibody (1:5000, 80713-1-RR; Proteintech), Anti-IRG1 antibody (1:1000; ab222411; abcam; Cambridge, UK), Nrf2 monoclonal antibody (1:2000; A21176; abclonal; Wuhan, China), iNOS Polyclonal antibody (1:500; 22226-1-AP; Proteintech), NOX2 Polyclonal antibody (1:3000; 19013-1-AP; Proteintech), p-p38 MAPK Polyclonal antibody (1:2000; 28796-1-AP; Proteintech), and p38 MAPK Polyclonal antibody (1:4000; 14064-1-AP; Proteintech).

Techniques: Derivative Assay, Fluorescence, Expressing, Cell Culture, Flow Cytometry, Western Blot, Control

Journal: Biomarker research

Article Title: S100A9 as a potential novel target for experimental autoimmune cystitis and interstitial cystitis/bladder pain syndrome.

doi: 10.1186/s40364-025-00763-5

Figure Lengend Snippet: Fig. 7 S100A9 promotes macrophage activation and secretion of inflammatory factors. (A) LPS promoted S100A9 secretion in mice primary macro phages (n = 6). (B-C) S100A9 promoted pro-inflammatory polarisation of primary mouse macrophages (n = 3). (D) S100A9 promoted IL-6, TNF-α, and IL-1β expression in mouse primary macrophages (n = 6). (E) Paquinimod inhibited the increased IL-6, TNF-α and IL-1β mRNA expression in S100A9-induced mouse macrophages (n = 9). (F) Paquinimod inhibited the increase of IL-6, TNF-α and IL-1β levels in S100A9-induced mouse macrophages (n = 9). (G) S100A9 activated TLR4/NF-κB and TLR4/p38 signalling pathways in primary mouse macrophages (n = 3). NS indicates no difference; *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The antibodies used in this study were as follows: rabbit monoclonal anti-S100A9 antibody (1:800, 73425, CST, USA), mouse monoclonal anti-TLR4 antibody (1:1000, MA5-16216, Thermo Fisher, USA), mouse monoclonal anti-MyD88 antibody (1:1000, 67969-1-Ig, Proteintech, China), rabbit polyclonal anti-P38 (1:1000, B-IO-10130, Biozellen, USA), rabbit polyclonal p-p38 (1:800, B-IO10131, Biozellen, USA), rabbit monoclonal anti-NF-κB (1:1000, 8242, CST, USA), rabbit monoclonal anti-pNF-κB (1:800, 3033, CST, USA), mouse monoclonal anti-IκBα antibody (1:1000, 4814, CST, USA), mouse monoclonal anti-IκBα antibody (1:1000, 4814, CST, USA), rabbit monoclonal anti-IκBα antibody (1:1000, 2859, CST, USA), mouse monoclonal anti-IL-1β (1:800, sc-12742, Santa Cruz Biotechnology, USA), mouse monoclonal anti-IL-6 (1:500, sc-57315, Santa Cruz Biotechnology, USA), TNF-α (1:1000, 17590-1-AP, Proteintech, China), rabbit polyclonal anti-TNF-α (1:600, 17590-1-AP, Proteintech, China), mouse monoclonal anti-GAPDH (1:1000, 60004-1-Ig, Proteintech, China), mouse monoclonal anti-caspase-3 (1:600, 66470-2-Ig, Proteintech, China), rabbit polyclonal anti-caspase-8 (1:600, 13423-1-AP, Proteintech, China), rabbit monoclonal anti-caspase-1 antibody (1:800, 24232, CST, USA), mouse monoclonal anti-Bax (1:1000, 60267-1-Ig, Proteintech, China), mouse monoclonal anti-UPK3A (1:800, sc-166808, Santa Cruz Biotechnology, USA), rabbit polyclonal anti-UPK2 (1:600, 21149-1-AP, Proteintech, China), goat anti-rabbit secondary antibody (1:2000, G6120, Thermo Fisher, USA) and goat anti-mouse secondary antibody (1:2000, G21040, Thermo Fisher, USA).

Techniques: Activation Assay, Expressing

Journal: Biomarker research

Article Title: S100A9 as a potential novel target for experimental autoimmune cystitis and interstitial cystitis/bladder pain syndrome.

doi: 10.1186/s40364-025-00763-5

Figure Lengend Snippet: Fig. 9 Knockdown of S100A9 significantly reduced TLR4/NF-κB and TLR4/p38 signalling pathway activating and decreasing inflammation and apoptosis- related protein expression in EAC mice (n = 6). GSEA analysis showing that TLR4/MyD88, NF-κB, and p38 signalling are activated in IC/BPS patients and EAC mice. (B-C) Knockdown of S100A9 significantly reduced TLR4/NF-κB and TLR4/p38 signalling pathway protein expression in EAC mice. (D-E) Knockdown of S100A9 significantly reduced the expression of bladder inflammation-related proteins (IL-6, IL-1β and TNF-α) in EAC mice. (F) GSEA analysis showing that apoptosis signalling is activated in IC/BPS patients and EAC mice. (G-H) Knockdown of S100A9 expression significantly reduced the expression of bladder apoptosis-related proteins (Bax, caspase-3, caspase-8, and caspase-1) and improved the expression of epithelial damage marker proteins (UPK3A and UPK2) in EAC mice. NS indicates no difference; *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: The antibodies used in this study were as follows: rabbit monoclonal anti-S100A9 antibody (1:800, 73425, CST, USA), mouse monoclonal anti-TLR4 antibody (1:1000, MA5-16216, Thermo Fisher, USA), mouse monoclonal anti-MyD88 antibody (1:1000, 67969-1-Ig, Proteintech, China), rabbit polyclonal anti-P38 (1:1000, B-IO-10130, Biozellen, USA), rabbit polyclonal p-p38 (1:800, B-IO10131, Biozellen, USA), rabbit monoclonal anti-NF-κB (1:1000, 8242, CST, USA), rabbit monoclonal anti-pNF-κB (1:800, 3033, CST, USA), mouse monoclonal anti-IκBα antibody (1:1000, 4814, CST, USA), mouse monoclonal anti-IκBα antibody (1:1000, 4814, CST, USA), rabbit monoclonal anti-IκBα antibody (1:1000, 2859, CST, USA), mouse monoclonal anti-IL-1β (1:800, sc-12742, Santa Cruz Biotechnology, USA), mouse monoclonal anti-IL-6 (1:500, sc-57315, Santa Cruz Biotechnology, USA), TNF-α (1:1000, 17590-1-AP, Proteintech, China), rabbit polyclonal anti-TNF-α (1:600, 17590-1-AP, Proteintech, China), mouse monoclonal anti-GAPDH (1:1000, 60004-1-Ig, Proteintech, China), mouse monoclonal anti-caspase-3 (1:600, 66470-2-Ig, Proteintech, China), rabbit polyclonal anti-caspase-8 (1:600, 13423-1-AP, Proteintech, China), rabbit monoclonal anti-caspase-1 antibody (1:800, 24232, CST, USA), mouse monoclonal anti-Bax (1:1000, 60267-1-Ig, Proteintech, China), mouse monoclonal anti-UPK3A (1:800, sc-166808, Santa Cruz Biotechnology, USA), rabbit polyclonal anti-UPK2 (1:600, 21149-1-AP, Proteintech, China), goat anti-rabbit secondary antibody (1:2000, G6120, Thermo Fisher, USA) and goat anti-mouse secondary antibody (1:2000, G21040, Thermo Fisher, USA).

Techniques: Knockdown, Expressing, Marker

Journal: Chinese Medical Journal

Article Title: S100A9 as a promising therapeutic target for diabetic foot ulcers

doi: 10.1097/CM9.0000000000003543

Figure Lengend Snippet: Inflammatory factors and epidermal thickness in chronic diabetic ulcers. (A, B) H&E staining of chronic ulcers of DM and Ctrl patients, and epidermal thickness (40× magnification, n = 3 per group). (C) Western blotting showing the levels of inflammatory factors (TNF-α, IL-1, and IL-6) in chronic ulcers of DM and Ctrl patients. GAPDH was used as an internal reference ( n = 3 per group). (D) Western blots showing the levels of signaling pathway proteins (p-ERK, p-p38, p-p65, p-Akt) in chronic ulcers of DM and Ctrl patients ( n = 3 per group). GAPDH was used as an internal reference. Akt: Protein kinase B; Ctrl: Non-diabetic group; DM: Diabetic group; ERK: Extracellular signal-regulated kinase; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; H&E: Hematoxylin and eosin; IL: Interleukin; P38: Protein 38; P65: Protein 65; p-: Phosphorylated; TNF-α: Tumor necrosis factor-α.

Article Snippet: The membranes were incubated with the following primary antibodies: S100A9 polyclonal antibody (CAT#14226-1-AP, Proteintech, Wuhan, China, 1:500), β-actin polyclonal antibody (CAT#20536-1-AP, Proteintech, Wuhan, China, 1:2000), GAPDH monoclonal antibody (CAT#60004-1-Ig, Proteintech, Wuhan, China, 1:10,000), phosphorylated (p)-protein kinase B (Akt) polyclonal antibody (CAT#28731-1-AP; Proteintech, Wuhan, China, 1:2000), interleukin (IL)-1α polyclonal antibody (CAT#DF6893, Affinity, Massachusetts, USA, 1:1000), IL-6 polyclonal antibody (CAT#DF6087, Affinity, Massachusetts, USA, 1:1000), tumor necrosis factor-α (TNF-α) polyclonal antibody (CAT#DF6080, Affinity, Massachusetts, USA, 1:1000), anti-p-Erk1+Erk2 recombinant rabbit monoclonal antibody (CAT#ET1610-13, Huabio, Zhejiang, China, 1:5000), p-nuclear factor-κB (NF-κB) p65 rabbit monoclonal antibody (CAT#3033, CST, Massachusetts, USA, 1:1000), and p-p38 mitogen-activated protein kinase (MAPK) rabbit polyclonal antibody (CAT#AP0526, ABclonal, Wuhan, China, 1:5000).

Techniques: Staining, Western Blot

Journal: Aging cell

Article Title: Negative Effect of Gst-35 on the Health Span of Caenorhabditis elegans Through Lysosomal Dysfunction via the Pmk-1 and Skr Genes.

doi: 10.1111/acel.70016

Figure Lengend Snippet: FIGURE 5 | Gst-35oe impacts the health span of nematodes by stimulating the PMK-1 pathway. (A) The survival curves of WT and OE nematodes fed either vector L4440 control bacteria or bacteria expressing RNAi for pmk-1. (B–E) The body length, body area, and moving speed of WT and OE nematodes fed vector L4440 control bacteria or bacteria expressing RNAi for pmk-1 on Day 1. (F, G) Western blotting of p-p38 mitogen–activated pro- tein kinase (MAPK) protein expression levels in WT, KO, and OE nematodes. (H) Representative fluorescence images of WT, OE, and gst-35oe/skr- 10oe nematodes (treated with vector RNAi or pmk-1 RNAi) stained with LysoTracker Red on Day 1 and its quantitative determination of acidic fluorescence intensity in the intestine. Data were represented as mean ± SEM of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 vs. OE with vector RNAi.

Article Snippet: Protein samples were run at 30 V for 40 min on a stacking gel and at 80 V for 120 min on a separating gel. p- p38 MAPK (Thr180/Tyr182) polyclonal antibody (dilution 1:1000, Proteintech) was used to determine the p- p38 levels.

Techniques: Plasmid Preparation, Control, Bacteria, Expressing, Western Blot, Fluorescence, Staining