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: Free radical biology & medicine

Article Title: N-Alkyl triphenylvinylpyridinium conjugated dihydroartemisinin perturbs mitochondrial functions resulting in enhanced cancer versus normal cell toxicity

doi: 10.1016/j.freeradbiomed.2021.01.050

Figure Lengend Snippet: TPVP-DHA treatment of MIA PaCa-2 cells significantly decreases phosphorylation status of MAPK signaling pathways. MIA PaCa-2 cells were treated with 5 μM of TPVP-DHA for 72 h. Two hundred micrograms of total protein extracts were incubated with RayBiotec MAPK Phosphorylation Arrays following the manufacturer supplied protocol. Chemiluminescence was used to image the membrane and results were quantified using NIH ImageJ software. Images of the original blot and heat maps are shown on left and quantitative results are shown on right. Percent change in MAPK-phosphorylation status was calculated relative to MAPK-phosphorylation in cells treated with 6-TPVP alone.

Article Snippet: Cell growth and cell cycle progression were measured using the Gen5™ software as described by the manufacturer. . RayBiotec MAPK Pathway arrays: Asynchronous cultures were treated with 5 μM DHA, 6-TPVP and TPVP-DHA for 72 h. Total protein extracts were prepared following the manufacturer supplied protocol and two hundred microgram of total protein extracts were incubated with RayBiotec MAPK Pathway Phosphorylation Arrays.

Techniques: Incubation, Software

Journal: Current Protocols in Protein Science

Article Title: Overview of Protein Microarrays

doi: 10.1002/0471140864.ps2701s72

Figure Lengend Snippet: Commercially Available Protein Microarrays

Article Snippet: Antibody for pathway detection , MAPK Pathway Phospho Antibody Array , BioCat , Analytical , 185 antibodies against phospho‐proteins in the MAPK pathways.

Techniques: Functional Assay, Microarray, Phospho-proteomics, Ab Array, Clinical Proteomics

Journal: Molecular Oncology

Article Title: KLK6‐regulated miRNA networks activate oncogenic pathways in breast cancer subtypes

doi: 10.1016/j.molonc.2016.03.008

Figure Lengend Snippet: The effect of KLK6 transfection on miRNA‐mediated signaling pathways. Panels of MAPK signaling and related factors were assayed for expression changes between KLK6 overexpressing and mock cells. In general, expression of mRNAs encoding for transcription ...

Article Snippet: The cDNA was synthesized as previously and was added to the MAPK Signaling Pathway RT 2 ProfilerTM PCR array (Qiagen/SuperArray Biosciences) as per the manufacturer's protocol.

Techniques: Transfection, Protein-Protein interactions, Expressing