Journal: Disease Markers

Article Title: Aldolase A Promotes Colorectal Cancer Progression through Targeting COPS6 and Regulating MAPK Signaling Pathway

doi: 10.1155/2023/1702125

Figure Lengend Snippet: Graphical abstract. By binding to and targeting COPS6, aberrantly expressed ALDOA promoted the EMT process and activated the ERK1/2 and P38 signaling pathways, ultimately accelerating CRC cell proliferation and metastasis.

Article Snippet: The following antibodies were used: ALDOA (mouse monoclonal; cat. no. sc-390733; 1 : 1000 dilution; Santa Cruz, CA, USA), HRP-conjugated DYKDDDDK Tag (monoclonal; cat. no. HRP-66008; 1 : 5000 dilution; Proteintech Group), ALDOB (rabbit polyclonal; cat. no. 18065-1-AP; 1 : 1000 dilution; Proteintech Group), ALDOC (rabbit polyclonal; cat. no. 14884-1-AP; 1 : 1000 dilution; Proteintech Group), E-cadherin (rabbit monoclonal; cat. no. 3195; 1 : 1000 dilution; Cell Signaling Technology), N-cadherin (rabbit monoclonal; cat. no. 13116; 1 : 1000 dilution; Cell Signaling Technology), vimentin (rabbit monoclonal; cat. no. 5741; 1 : 1000 dilution; Cell Signaling Technology), p38 (rabbit monoclonal; cat. no. 8690; 1 : 1000 dilution; Cell Signaling Technology), p-p38 (rabbit monoclonal; cat. no. 8632; 1 : 1000 dilution; Cell Signaling Technology), ERK1/2 (rabbit monoclonal; cat. no. 4695; 1 : 1000 dilution; Cell Signaling Technology), p-ERK1/2 (rabbit monoclonal; cat. no. 4376; 1 : 1000 dilution; Cell Signaling Technology), ACTB (rabbit monoclonal; cat. no. AC038; 1 : 10000 dilution; ABclonal Technology, Wuhan, China), GAPDH (rabbit monoclonal; cat. no. 60004-1-Ig; 1 : 10000 dilution; Proteintech Group), lamin B1 (rabbit polyclonal; cat. no. 12987-1-AP; 1 : 2000 dilution; Proteintech Group), PKM (rabbit polyclonal; cat. no. 10078-2-AP; 1 : 1000 dilution; Proteintech Group), HSP90AB (rabbit polyclonal; cat. no. RK05737; 1 : 1000 dilution; ABclonal Technology), CSN6 (mouse monoclonal; cat. no. sc-393023; 1 : 1000 dilution; Santa Cruz, CA, USA), caspase-3 (rabbit monoclonal; cat. no. 9662; 1 : 1000 dilution; Cell Signaling Technology), and cleaved caspase-3 (rabbit monoclonal; cat. no. 9661; 1 : 1000 dilution; Cell Signaling Technology).

Techniques: Binding Assay, Protein-Protein interactions

Journal: Frontiers in oncology

Article Title: CDKN2A Deletion in Melanoma Excludes T Cell Infiltration by Repressing Chemokine Expression in a Cell Cycle-Dependent Manner.

doi: 10.3389/fonc.2021.641077

Figure Lengend Snippet: FIGURE 3 | CDKN2A enhances chemokine expression. (A) Chemokine expression in CDKN2A Diploid and Deep deletion melanoma cell lines. (B) Cell cycle analysis of vehicle, double thymidine treated B16F0 cells and CDKN2A re-expressed B16F0 cells. (C) The mRNA expression of chemokine Ccl4, Ccl5, Cxcl9, Cxcl10, and Cxcl11 expression in control, CDKN2A re-expressed and double thymidine treated B16F0 cells. (D) ELISA analysis of the chemokine Ccl4, Ccl5, and Cxcl10 expression in control, CDKN2A re-expressed and double thymidine treated B16F0 cells. (E) The mRNA expression of chemokine Ccl4, Ccl5, Cxcl9, Cxcl10, and Cxcl10 in B16F0 cells arrested at G1/S and the cells arrested at G2/M phase. (F) The GSEA analysis of the gene expression in P38/MAPK and NF-kB pathway in patients with normal CDKN2A or CDKN2A deletion in TCGA-SKCM cohort. (G) The expression of protein in p38/MAPK and NF-kB pathway in B16F0 cells arrested at G1/S phase, S phase, G2/M phase with or without H2O2 treatment. (H) mRNA expression of chemokine Ccl4, Ccl5, Cxcl9, Cxcl10, and Cxcl11 in B16F0 cells treated with H2O2 or not. (I) The mRNA expression of genes downstream of p38/MAPK and NF-kB pathway in B16F0 cells arrested at G1/S phase, S phase, G2/M phase with or without H2O2 treatment. *p < 0.05, **p < 0.01, ***P < 0.001, ns, not significant.

Article Snippet: The membrane was blocked with 5% nonfat milk in TBST and incubated with a primary antibody against Vinculin (1:1000, sc-73264, RRID : AB_1131292, Santa Cruz, USA), phospho-p65 (Ser536) (1:1000, #3031S, RRID : AB_330559, Cell Signaling Technology, Boston, USA), phospho-p38 MAPK (Thr180/Tyrl82) (1:1000, #4631S, RRID : AB_331765, Cell Signaling Technology), phospho-ATF2 (T71) (1:1000, BS4018, RRID : AB_1664091, Bioworld, China), phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:1000, #4370P, RRID : AB_2315112, Cell Signaling Technology), and IkBa (1:1000, #9242S, RRID : AB_331623, Cell Signaling Technology), CDK4 (1:500, sc-23896, RRID : AB_627239, Santa Cruz), E2F1 (1:1000, 12171-1-AP, RRID : AB_2096958, Proteintech), p16INK4 (1:500, sc-1661, RRID : AB_628067, Santa Cruz), Phospho-Histone H3 (1:1000, #9701, RRID : AB_331535, Cell Signaling Technology), Cyclin D1 (1:200, sc753, RRID : AB_2070433, Santa Cruz), Cyclin E1 (1:500, BS1086, RRID : AB_1663604, Bioworld), Cyclin B1 (1:1000, sc-245, RRID ;: AB_627338, Santa Cruz), at 4°C overnight.

Techniques: Expressing, Cell Cycle Assay, Control, Enzyme-linked Immunosorbent Assay, Gene Expression

Journal: eLife

Article Title: p38γ and p38δ modulate innate immune response by regulating MEF2D activation

doi: 10.7554/eLife.86200

Figure Lengend Snippet: ( A ) Samples of genomic DNA from wild-type (WT), Mapk13 −/− , Mapk12 171A/171A , and p38γ/δKIKO purified from tail biopsy were used as templates for PCR as described in methods. ( B ) WT and p38γ/δKIKO mouse embryonic fibroblasts (MEFs) were exposed to 0.5 M sorbitol (15 min), activation and expression of MAPK were examined in immunoblotted lysates (30 µg) using phospho-specific and specific antibodies for each indicated protein. ( C ) MEFs were treated as in ( B ); endogenous p38γ immunoprecipitated with anti-p38γ from 2 mg cell lysate and immunoblotted with the p38 phospho-specific antibody, which recognizes all phosphorylated p38 isoforms. ( D ) Endogenous hDlg was immunoprecipitated with anti-hDlg from 1 mg MEF lysate (treated as in ( B )). Pellets were immunoblotted with the indicated antibodies. Red (*) mark unspecific protein bands. ( E ) Extracts from WT, p38γ/δKO, and p38γ/δKIKO peritoneal macrophages were immunoblotted using specific antibodies for TPL2 and p38α. Representative blots of two independent experiments are shown in all panels. Figure 1—figure supplement 1—source data 1. Labelled (.pdf) and raw (.tif) western blot images showed in panel A. Figure 1—figure supplement 1—source data 2. Labelled (.pdf) and raw (.tif,. jpeg) western blot images showed in panel B. Figure 1—figure supplement 1—source data 3. Labelled (.pdf) and raw (.jpeg) western blot images showed in panel C. Figure 1—figure supplement 1—source data 4. Labelled (.pdf) and raw (.jpeg) western blot images showed in panel D. Figure 1—figure supplement 1—source data 5. Labelled (.pdf) and raw (.jpeg) western blot images showed in panel E.

Article Snippet: P-p38 , Phospho-p38 MAP Kinase (Thr180/Tyr182) Antibody , Cell Signaling , WB , 1/1000.

Techniques: Purification, Activation Assay, Expressing, Immunoprecipitation, Western Blot

Journal: eLife

Article Title: p38γ and p38δ modulate innate immune response by regulating MEF2D activation

doi: 10.7554/eLife.86200

Figure Lengend Snippet: Information about the antibodies used in this study.

Article Snippet: P-p38 , Phospho-p38 MAP Kinase (Thr180/Tyr182) Antibody , Cell Signaling , WB , 1/1000.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Research on Roles of Mongolian Medical Warm Acupuncture in Inhibiting p38 MAPK Activation and Apoptosis of Nucleus Pulposus Cells

doi: 10.1155/2018/6571320

Figure Lengend Snippet: Phosphorylation level of p38 MAPK in the nucleus pulposus cells and effect of acupuncture on substrate metabolism of the rat intervertebral disc. (a) The phosphorylation level of p38 MAPK decreased following administration of analgesic decoction. (b/c) The expression of the Type II collagen, aggrecan, Sox-9, and TIMP-1 in the nucleus pulposus cells in the model group significantly decreased when compared with that in the control group. The expression of MMP-3 increased significantly when compared with that in the control group. # (##) indicates P<0.05 (P<0.01) when compared with the control group; ∗ ( ∗∗ ) indicates P<0.05 (P<0.01) when compared with the 25 mM glucose group.

Article Snippet: The protein sample was transferred to the nitrocellulose membrane, blocked, and incubated with the p38 MAPK antibody (dilution rate 1 : 1 000) (Miltenyi Biotec, Bergisch-Gladbach, Germany) and the GAPDH antibody (1 : 2 000) (Wuhan Boster Biological Engineering Co., Ltd., Wuhan, China), respectively, for 2 h at room temperature, incubated with the appropriate secondary antibody (Wuhan Boster Biological Engineering Co., Ltd., Wuhan, China) for 1 h following membrane washing, luminized, and developed with the chemiluminescence method.

Techniques: Phospho-proteomics, Expressing, Control