Journal: Oral oncology

Article Title: P2Y 2 receptors mediate nucleotide-induced EGFR phosphorylation and stimulate proliferation and tumorigenesis of head and neck squamous cell carcinoma cell lines

doi: 10.1016/j.oraloncology.2020.104808

Figure Lengend Snippet: Selective inhibition of P2Y2R, ADAM10/17, EGFR and Src kinase diminishes UTP-induced EGFR phosphorylation. Representative immunoblots of CAL27 cells pretreated for 1 h with or without (A) AR-C118925 (10 μM), (B) TAPI-2 (10 μM), (C) cetuximab (10 μg/ml) or (D) dasatinib (100 nM) followed by 100 μM UTP or 10 ng/ml EGF for 1 min, then subjected to immunoblot analysis using antibodies for phospho-EGFR (Tyr1173) or total EGFR as a loading control. (E) UTP- or (F) EGF-induced EGFR (Tyr1173) phosphorylation levels were quantified and data are presented as the percentage of maximal EGFR phosphorylation induced by UTP or EGF alone. Data are presented as means ± S.E.M. for at least n = 3 experiments, where ** indicates P < 0.01, *** indicates P < 0.001 and **** indicates P < 0.0001.

Article Snippet: AR-C118925, AG1478, {"type":"entrez-nucleotide","attrs":{"text":"GF109203","term_id":"295317075","term_text":"GF109203"}} GF109203 and TAPI-2 were purchased from Bio-Techne Corporation (Minneapolis, MN).

Techniques: Inhibition, Western Blot

Journal: iScience

Article Title: Heightened innate immune state induced by viral vector leads to enhanced response to challenge and prolongs malaria vaccine protection

doi: 10.1016/j.isci.2024.111468

Figure Lengend Snippet:

Article Snippet: TAPI2 , Biosynth , Cat #INH-3852-PI.

Techniques: Control, Virus, Recombinant, Giemsa Stain, Cell Stimulation, Transfection, Software

Journal: Cell Death and Differentiation

Article Title: ADAM12-directed ectodomain shedding of E-cadherin potentiates trophoblast fusion

doi: 10.1038/cdd.2015.44

Figure Lengend Snippet: ADAM12S controls trophoblast fusion in part by E-cadherin ectodomain shedding. (a) Representative immunoblots (N=3) of CM or whole cell lysates (WCL) from ADAM12S-transfected Bewo cells cultured for 72 h in the absence (−) or presence of the broad-spectrum metalloproteinase inhibitors, TAPI-1 (T1) or TAPI-2 (T2). Lysates were probed with antibodies directed against ADAM12 (pro-A12S; mature-A12S) or E-cadherin (Ecad). Molecular weights (kDa) are shown to the left and β-actin indicate loading controls. (b) Representative immunofluorescence images (N=3) of ADAM12S-transfected Bewo cells probed with anti-E-cadherin antibody (Ecad; red). Cells were either untreated (−) or cultured in the presence of TAPI-2; DAPI-stained nuclei are labeled blue. White perforated box indicates magnified area. (c) ADAM12 (green) and E-cadherin (Ecad; red) immunofluorescence images of Bewo cells stably transfected with either wildtype ADAM12S or protease-dead ADAM12SΔE351Q expression constructs; Bars=50 μm. (d) Immunoblots (N=3) of ADAM12S and E-cadherin in CM or WCL of Bewo cells stably transfected with the control pCMV6 construct as well as expression constructs described in (c). Molecular weights (kDa) are shown to the left and β-actin indicates loading control. (e) Box-plots depicting median and IQR values of the percent of multi-nucleated cells in ADAM12S and ADAM12SΔE351Q Bewo cells (N=3) cultured over 72 h; *P≤0.05. (f) Representative immunoblots (N=3) probed with antibodies directed against E-cadherin (Ecad), GFP and ADAM12 showing ADAM12-directed E-cadherin cleavage. Immunoprecipitated ADAM12-GFP was incubated with recombinant GST N-tagged E-cadherin (GST-rEcad); immunoprecipitated GFP and ADAM12ΔE351Q-GFP serve as controls. The full-length (fl) and cleaved (cl) E-cadherin products are indicated; ns denotes non-specific bands. Molecular weights (kDa) are shown to the left

Article Snippet: The broad-spectrum metalloproteinase inhibitors, TAPI-1 (10 μ M; Enzo Life Sciences, Farmingdale, NY, USA) and TAPI-2 (10 μ M; Enzo Life Sciences), were added to culture media in select experiments; 0.5% DMSO was added to control cells.

Techniques: Western Blot, Transfection, Cell Culture, Immunofluorescence, Staining, Labeling, Stable Transfection, Expressing, Construct, Immunoprecipitation, Incubation, Recombinant