Journal: Infection and Immunity

Article Title: Thioredoxin from Brugia malayi: Defining a 16-Kilodalton Class of Thioredoxins from Nematodes

doi: 10.1128/IAI.71.7.4119-4126.2003

Figure Lengend Snippet: Bm-TRX-1 inhibits activation of p38 MAP kinase. The WCPPCR and WCPQCR alleles of Bm-trx-1 were cloned separately into a eukaryotic expression plasmid vector, and the plasmids were introduced into TPA-differentiated ML-1 cells. The transfected ML-1 cells were stimulated with TNF-α (100 IU/well) for 5 min, and the phosphorylation status of p38 MAP kinase was analyzed on immunostained Western blots of cell extracts. Controls included cells maintained in medium (medium control), cells exposed to the transfection regent (transfection control), and cells transfected with a plasmid containing no insert (plasmid control).

Article Snippet: In preliminary experiments, TPA-differentiated ML-1 cells were stimulated with TNF-α (100 IU/well) for 5, 10, and 30 min, and the phosphorylation status of p38 MAP kinase was analyzed on immunostained Western blots of cell extracts with phosphoPlus p38 MAP kinase antibody kit (Cell Signaling Technology, Beverly, Mass.).

Techniques: Activation Assay, Clone Assay, Expressing, Plasmid Preparation, Transfection, Western Blot

Journal: Nature

Article Title: Neutrophil ageing is regulated by the microbiome

doi: 10.1038/nature15367

Figure Lengend Snippet: Pathways selected for the analysis of neutrophil functions.

Article Snippet: MAPK pathway , BIOCARTA_MAPK_PATHWAY.

Techniques: Coagulation, Activation Assay, Ubiquitin Proteomics

Journal: Nature

Article Title: Neutrophil ageing is regulated by the microbiome

doi: 10.1038/nature15367

Figure Lengend Snippet: Gene set enrichment analysis of selected pathways in aged and activated neutrophils.

Article Snippet: MAPK pathway , BIOCARTA_MAPK_PATHWAY.

Techniques: Coagulation, Activation Assay, Ubiquitin Proteomics

Journal: ACR Open Rheumatology

Article Title: Inhibition of Ras GTPases prevents Collagen‐Induced Arthritis by Reducing the Generation of Pathogenic CD4 + T Cells and the Hyposialylation of Autoantibodies

doi: 10.1002/acr2.11169

Figure Lengend Snippet: Farnesylthiosalicylate (FTS) inhibits phosphorylation of mitogen‐activated protein kinases (MAPKs) and T‐cell proliferation following T‐cell antigen receptor (TCR) stimulation. Isolated mouse CD4+ T cells were stimulated with anti‐‐CD3/28 antibodies and treated with FTS 10 µM (green) or vehicle (red) for 30 minutes. A , Bar graphs depict the relative phosphorylation level (arbitrary units of intensity per densitometry) of the indicated kinases included in the RayBio Mouse MAPK Pathway Phosphorylation Array, as analyzed per the manufacturer’s protocol. B , Flow cytometry data expressed as overlay histograms depicting phosphorylated (p)‐extracellular signal‐regulated kinase (Erk)1/2, p‐AKT, and p‐p38 levels, determined by phospho‐flow‐cytometry with phospho‐specific antibodies, as detailed in the Methods section. T cells treated with FTS or control media are shown in green and red, respectively, whereas unstained counterpart samples are in grey. C , CD4+ T cells were labeled with carboxyfluorescein succinimidyl ester (CFSE) and activated by plate‐bound anti‐CD3 and soluble anti‐CD28 monoclonol antibodies (mAbs), treated with the indicated concentration of FTS, and cultured for 72 hours. Shown is a representative experiment (CFSE‐dilution flow cytometry histograms) out of two independent experiments, and the bar graphs depict mean ± SD of triplicates. Samples were analyzed for statistical significance by Student's t test (*** P < .001, ** P < .01, and * P < .05).

Article Snippet: Using the RayBio Mouse MAPK Pathway Phosphorylation Array, we determined the relative phosphorylation level of 17 different kinases, including well‐established Ras effectors (Figure ).

Techniques: Isolation, Flow Cytometry, Labeling, Concentration Assay, Cell Culture

Journal: BMC Cancer

Article Title: Involvement of the CB 2 cannabinoid receptor in cell growth inhibition and G0/G1 cell cycle arrest via the cannabinoid agonist WIN 55,212–2 in renal cell carcinoma

doi: 10.1186/s12885-018-4496-1

Figure Lengend Snippet: PI3K/Akt and MAPK/ERK1/2 pathways activation profile of RCC cells. a Dual pathway (PI3K and MAPK) profile of 786-O cells treated with agonist WIN-55 (0–25 μM). b Dual pathway (PI3K and MAPK) profile of ACHN cells treated with agonist WIN-55 (0–25 μM). ACHN and 786-O cells were found positive for Phospho-ERK1/2 (MAPK/ERK1/2) but negative for Phospho-Akt (PI3K/Akt). The dot plots showing WIN-55 treatment does not activate/inhibit PI3K and MAPK pathways even in higher concentrations (10–25 μM) as compared to the not treated control (0 μM, first column in Fig. a and b ). In each fig. a and b dot blots in second and third raw represents cells single stained with Phospho-Akt (PI3K/Akt) and Phospho-ERK1/2 (MAPK/ERK1/2), respectively. Dot blots in fourth raw in each fig. (a and b) represents cells double stained with Phospho-Akt (PI3K/Akt) and Phospho-ERK1/2 (MAPK/ERK1/2)

Article Snippet: The Muse™ PI3K/MAPK Dual Pathway Activation Kit (MCH200108) was purchased from Merck EMD Millipore (Massachusetts, USA) to assess the activation of PI3K and MAPK signaling pathways.

Techniques: Activation Assay, Staining