anti phospho p38 rabbit mab  (Cell Signaling Technology Inc)

Journal: Nature Communications

Article Title: Cholangiocarcinoma PDHA1 succinylation suppresses macrophage antigen presentation via alpha-ketoglutaric acid accumulation

doi: 10.1038/s41467-025-58429-7

Figure Lengend Snippet: a Gene Ontology (GO) analysis of RNA-seq after 24 h co-culture of tumor CM with or without α-KG (8 μM) and THP-1 cells. b Volcano plot to depict the downregulated HLA-DRA , HLA-DPA1 , and HLA-DQA1 expression changes of immune response observed in the RNA-seq data. c Gene Set Enrichment Analysis (GSEA) indicating that positive regulation of MAPK cascade was significantly upregulated after α-KG accumulation. d Western blot analysis showed the expression of the MAPK pathway and HLA-DR, CD74 changes after 24 h co-culture ( n = 3 biological replicates). The samples derive from the same experiment but different gels for p-ERK, ERK, and HLA-DR, another for CD74 and p-P38, another for p-JNK, JNK, and another for P38 and α-Tubulin were processed in parallel. e Diagram of interaction between α-KG and OXGR1 protein using molecular docking. f Western blot analysis showed the expression of p-ERK, HLA-DR, and CD74 changes after 24 h co-culture ( n = 3 biological replicates). The samples derive from the same experiment but different gels for p-ERK, ERK, and HLA-DR, another for CD74, and another for OXGR1 and α-Tubulin were processed in parallel. g Schematic diagram of THP-1 with or without OXGR1 depletion and tumor CM with α-KG (0, 8 μM). Created in BioRender. Zhang, N. (2025) https://BioRender.com/a10o519 . After 24 h, macrophages were used for flow analysis. h Flow cytometry analysis of HLA-DR + MFI values from data ( g ) ( n = 4 biological replicates). i Schematic diagram of subcutaneous tumor treatment. Created in BioRender. Zhang, N. (2025) https://BioRender.com/j80q160 . And representative images of vehicle, montelukast, α-KG or combined on day 26 ( n = 5 biologically independent samples). j Tumor growth of LTP-C9 with overexpression of PDHA1 WT were injected into C57BL/6 mice ( n = 5 biologically independent samples), with vehicle, montelukast, α-KG or combined treatment, beginning on day 14. k Tumor volume from data ( i ) ( n = 5 biologically independent samples). l Tumor weight from data ( i ) ( n = 5 biologically independent samples). Data are presented as mean values ± SD, and P values were calculated using two-tailed unpaired Student’s t -tests ( d , f , h , k , l ) or 2way ANOVA ( j ). Each Western blot was independently repeated three times. Mac macrophages, CM conditioned medium, MFI mean fluorescence intensity. Source data are provided as a Source Data File.

Article Snippet: The antibodies were as follows: anti-Succinyllysine Mouse mAb (419, 1:1000 for Western blot), anti-Succinyllysine Rabbit pAb (401, 1:1000 for Western blot), anti-Succinyl-PDHA1(K83) Rabbit pAb (Cat#CM0401, Lot#ZC492M726P1P-ET2, 1:1000 for Western blot, 1:200 for IHC, IF and mIHC) are from PTM BioLab, Co. Ltd. Anti-DYKDDDDK Tag Rabbit mAb (14793, 1:1000 for Western blot), anti-α-Tubulin (2144, 1:1000 for Western blot), anti-CD86 Rabbit mAb (91882, 1:1000 for Western blot, 1:200 for mIHC), anti-DLST Rabbit mAb (11954, 1:1000 for Western blot, 1:500 for IHC and 1:200 for IF), anti-CD206/MRC1 Rabbit mAb (24595, 1:1000 for Western blot, 1:200 for IHC), anti-β-Actin Antibody (4967, 1:1000 for Western blot), anti-Phospho-Erk1/2 Rabbit mAb (4370, 1:1000 for Western blot), anti-MAPK Rabbit mAb (4695, 1:1000 for Western blot), anti-Phospho-p38 Rabbit mAb (4511, 1:1000 for Western blot), anti-p38 MAPK Rabbit mAb (8690, 1:1000 for Western blot), anti-Phospho-SAPK/JNK Rabbit mAb (4668, 1:1000 for Western blot), anti-SAPK/JNK (9252, 1:1000 for Western blot), anti-Phospho-STAT1 Rabbit mAb (9167, 1:1000 for Western blot), anti-Phospho-NF-kB Rabbit mAb (3033, 1:1000 for Western blot), anti-Phospho-AKT Rabbit mAb (4060, 1:1000 for Western blot), anti-HA-Tag Rabbit mAb (3724, 1:1000 for Western blot), anti-His rabbit mAb (2365, 1:1000 for Western blot), anti-GST rabbit mAb (2622, 1:1000 for Western blot) are from Cell Signaling Technology.

Techniques: RNA Sequencing, Co-Culture Assay, Expressing, Western Blot, Flow Cytometry, Over Expression, Injection, Two Tailed Test, Fluorescence

Journal: Molecular Medicine Reports

Article Title: Recombinant human collagen XVII protects skin basement membrane integrity by inhibiting the MAPK and Wnt signaling pathways

doi: 10.3892/mmr.2025.13465

Figure Lengend Snippet: Regulatory effect of RHCXVII on protein phosphorylation. (A) Phospho-antibody array analysis was performed to assess changes in phosphoprotein expression in HaCaT cells with and without RHCXVII treatment. (B) Kyoto Encyclopedia of Genes and Genomes pathway analysis of differentially phosphorylated proteins in RHCXVII-treated HaCaT cells compared with NC. Changes in expression levels of upregulated and downregulated phosphoproteins in (C) MAPK and (D) Wnt pathways in RHCXVII-treated HaCaT cells compared with NC group. RHCXVII, recombinant human collagen XVII; NC, negative control; diff phospho, differentially phosphorylated.

Article Snippet: Collagen type IV α1 chain (COL4A1; cat. no. ab214417), COL7A1 (cat. no. ab309143), laminin subunit β3 (LAMB3; cat. no. ab14509), integrin α6 (ITGA6; cat. no. ab181551), MMP2 (cat. no. ab97779) and vinculin (cat. no. ab129002) antibodies were purchased from Abcam. p38 MAPK (cat. no. 9212S), phospho-p38 MAPK (Thr180/Tyr182; D3F9) XP ® rabbit mAb (cat. no. 4511S), c-Jun (60A8) rabbit mAb (cat. no. 9165S) and phospho-c-Jun (Ser243; cat. no. 2994) antibodies were obtained from Cell Signaling Technology, Inc.

Techniques: Ab Array, Expressing, Recombinant, Negative Control

Journal: Molecular Medicine Reports

Article Title: Recombinant human collagen XVII protects skin basement membrane integrity by inhibiting the MAPK and Wnt signaling pathways

doi: 10.3892/mmr.2025.13465

Figure Lengend Snippet: RHCXVII inhibits phosphorylation of proteins in the MAPK and Wnt signaling pathways in HaCaT cells. (A) Representative western blotting. The protein expression levels of (B) p38 (phospho-Tyr182)/p38, (C) c-Jun (phospho-Ser243)/c-Jun and (D) MMP2. ## P<0.01 vs. BC; **P<0.01 vs. NC. RHCXVII, recombinant human collagen XVII; NC, negative control; TGF-β1, transforming growth factor β1; PC, positive control; BC, blank control; phospho, phosphorylated.

Article Snippet: Collagen type IV α1 chain (COL4A1; cat. no. ab214417), COL7A1 (cat. no. ab309143), laminin subunit β3 (LAMB3; cat. no. ab14509), integrin α6 (ITGA6; cat. no. ab181551), MMP2 (cat. no. ab97779) and vinculin (cat. no. ab129002) antibodies were purchased from Abcam. p38 MAPK (cat. no. 9212S), phospho-p38 MAPK (Thr180/Tyr182; D3F9) XP ® rabbit mAb (cat. no. 4511S), c-Jun (60A8) rabbit mAb (cat. no. 9165S) and phospho-c-Jun (Ser243; cat. no. 2994) antibodies were obtained from Cell Signaling Technology, Inc.

Techniques: Western Blot, Expressing, Recombinant, Negative Control, Positive Control, Control

Journal: Molecular Medicine Reports

Article Title: Recombinant human collagen XVII protects skin basement membrane integrity by inhibiting the MAPK and Wnt signaling pathways

doi: 10.3892/mmr.2025.13465

Figure Lengend Snippet: Key signaling pathways affected by RHCXVII. RHCXVII protects the skin basement membrane integrity by increasing expression of key extracellular matrix components such as collagen IV, collagen VII, laminin 332 and ITGA6 and decreasing MMP2 expression. RHCXVII decreases the phosphorylation of p38 and c-Jun in the MAPK and Wnt pathways. RHCXVII, recombinant human collagen XVII; ITGA6, integrin α6; p-, phosphorylated.

Article Snippet: Collagen type IV α1 chain (COL4A1; cat. no. ab214417), COL7A1 (cat. no. ab309143), laminin subunit β3 (LAMB3; cat. no. ab14509), integrin α6 (ITGA6; cat. no. ab181551), MMP2 (cat. no. ab97779) and vinculin (cat. no. ab129002) antibodies were purchased from Abcam. p38 MAPK (cat. no. 9212S), phospho-p38 MAPK (Thr180/Tyr182; D3F9) XP ® rabbit mAb (cat. no. 4511S), c-Jun (60A8) rabbit mAb (cat. no. 9165S) and phospho-c-Jun (Ser243; cat. no. 2994) antibodies were obtained from Cell Signaling Technology, Inc.

Techniques: Membrane, Expressing, Recombinant