Journal: BMC Microbiology

Article Title: Global secretome characterization of A549 human alveolar epithelial carcinoma cells during Mycoplasma pneumoniae infection

doi: 10.1186/1471-2180-14-27

Figure Lengend Snippet: Immune response and stress related proteins that were up-regulated by Mycoplasma pneumoniae infection

Article Snippet: The primary antibodies utilized included rabbit polyclonal anti-ADAM9 antibody (Cell Signaling Technolgoy, Beverly, MA, USA, diluted 1:1000), rabbit polyclonal anti-Gal1 antibody (Proteintech, Chicago, IL, diluted 1:1500), rabbit polyclonal anti-MIF antibody (Proteintech, diluted 1:2000), rabbit polyclonal anti-IL33 antibody (Proteintech, diluted 1:600), rabbit polyclonal anti-SERPINE1 antibody (Proteintech, diluted 1:800), rabbit polyclonal anti-IGFBP4 antibody (Millipore, diluted 1:1000), mouse monoclonal anti-β-actin antibody (Upstate, Lake Placid, NY, diluted 1:3000).

Techniques: Migration

Journal: PLoS ONE

Article Title: Regulation of Amyloid Precursor Protein Processing by Serotonin Signaling

doi: 10.1371/journal.pone.0087014

Figure Lengend Snippet: (A) SH-SY5Y cells, transfected with pEAK12-AP-APP and pcDNA3.1-5-HT 4d , were treated with 1 µM prucalopride or 5-HT (5-HT 4 receptor agonists) in the absence or presence of 80 µM GM6001 (metalloproteinases inhibitor) and secretion of sAPPα was analyzed via measuring SEAP. (B) SEAP levels were measured in supernatants of SH-SY5Y cells, co-transfected with pEAK12-AP-APP, pcDNA3.1-5-HT 4d and 3 nM siRNA for knock-down of ADAM9 (A9), ADAM10 (A10), ADAM17 (A17) and MMP9 and treated with 1 µM prucalopride. (C) Cell lysates of (B) were analyzed for protein expression of ADAM9, 10, 17 and MMP9 by western blotting. The ADAM10 and ADAM17 immature precursor proteins are indicated by an x, whereas the mature catalytically active forms are indicated by an xx for ADAM9, 17 and MMP9. The immature ADAM9 and the mature ADAM10 proteins were not visible. (D) Quantification of experiments in (C). Values shown are mean ± SEM of 6 individual wells and were normalized to vehicle control. * P <0.05, ** P <0.01, *** P <0.001, one-way ANOVA with Tukey-Kramer's post-hoc test.

Article Snippet: Following antibodies were purchased: CK2 α (H-286) from Santa Cruz, MMP-9 (G657) and ADAM9 (2099) from Cell Signaling, ADAM17 (T5442) and β-Actin (A5441) from Sigma-Aldrich.

Techniques: Transfection, Expressing, Western Blot

Journal: PLoS ONE

Article Title: Regulation of Amyloid Precursor Protein Processing by Serotonin Signaling

doi: 10.1371/journal.pone.0087014

Figure Lengend Snippet: (A) and (B) SH-SY5Y cells, co-transfected with pEAK12-AP-APP, pcDNA3.1-5-HT 4 and combinations of 3 nM siRNA for knock-down of ADAM9 (A9) and ADAM10 (A10), ADAM9 (A9) and ADAM17 (A17), ADAM10 (A10) and ADAM17 (A17) in (A) or ADAM9, 10 and 17 or ADAM9, 10, 17 and MMP9 in (B), were treated with 1 µM prucalopride (5-HT 4 receptor agonist) in the absence or presence of 80 µM GM6001 (metalloproteinases inhibitor) (B) and secretion of sAPPα was analyzed via measuring SEAP. (C) Cell lysates of experiments in (A) and (B) were analyzed for protein expression of ADAM9, 10, 17 and MMP9 by western blotting. ADAM10 and ADAM17 immature precursor proteins are indicated by an x, whereas the mature catalytically active forms are indicated by an xx for ADAM9, 17 and MMP9. The immature ADAM9 and the mature ADAM10 proteins were not visible. (D) Quantification of experiments in (C). Values shown are mean ± SEM of 6 individual wells and were normalized to vehicle control. ** P <0.01, *** P <0.001, one-way ANOVA with Tukey-Kramer's post-hoc test.

Article Snippet: Following antibodies were purchased: CK2 α (H-286) from Santa Cruz, MMP-9 (G657) and ADAM9 (2099) from Cell Signaling, ADAM17 (T5442) and β-Actin (A5441) from Sigma-Aldrich.

Techniques: Transfection, Expressing, Western Blot

Journal: PLoS ONE

Article Title: Regulation of Amyloid Precursor Protein Processing by Serotonin Signaling

doi: 10.1371/journal.pone.0087014

Figure Lengend Snippet: (A) SH-SY5Y cells, transfected with pEAK12-AP-APP and pcDNA3.1-5-HT 4d , were treated with 1 µM prucalopride or 5-HT (5-HT 4 receptor agonists) in the absence or presence of 80 µM GM6001 (metalloproteinases inhibitor) and secretion of sAPPα was analyzed via measuring SEAP. (B) SEAP levels were measured in supernatants of SH-SY5Y cells, co-transfected with pEAK12-AP-APP, pcDNA3.1-5-HT 4d and 3 nM siRNA for knock-down of ADAM9 (A9), ADAM10 (A10), ADAM17 (A17) and MMP9 and treated with 1 µM prucalopride. (C) Cell lysates of (B) were analyzed for protein expression of ADAM9, 10, 17 and MMP9 by western blotting. The ADAM10 and ADAM17 immature precursor proteins are indicated by an x, whereas the mature catalytically active forms are indicated by an xx for ADAM9, 17 and MMP9. The immature ADAM9 and the mature ADAM10 proteins were not visible. (D) Quantification of experiments in (C). Values shown are mean ± SEM of 6 individual wells and were normalized to vehicle control. * P <0.05, ** P <0.01, *** P <0.001, one-way ANOVA with Tukey-Kramer's post-hoc test.

Article Snippet: Following antibodies were purchased: CK2 α (H-286) from Santa Cruz, MMP-9 (G657) and ADAM9 (2099) from Cell Signaling, ADAM17 (T5442) and β-Actin (A5441) from Sigma-Aldrich.

Techniques: Transfection, Expressing, Western Blot

Journal: PLoS ONE

Article Title: Regulation of Amyloid Precursor Protein Processing by Serotonin Signaling

doi: 10.1371/journal.pone.0087014

Figure Lengend Snippet: (A) and (B) SH-SY5Y cells, co-transfected with pEAK12-AP-APP, pcDNA3.1-5-HT 4 and combinations of 3 nM siRNA for knock-down of ADAM9 (A9) and ADAM10 (A10), ADAM9 (A9) and ADAM17 (A17), ADAM10 (A10) and ADAM17 (A17) in (A) or ADAM9, 10 and 17 or ADAM9, 10, 17 and MMP9 in (B), were treated with 1 µM prucalopride (5-HT 4 receptor agonist) in the absence or presence of 80 µM GM6001 (metalloproteinases inhibitor) (B) and secretion of sAPPα was analyzed via measuring SEAP. (C) Cell lysates of experiments in (A) and (B) were analyzed for protein expression of ADAM9, 10, 17 and MMP9 by western blotting. ADAM10 and ADAM17 immature precursor proteins are indicated by an x, whereas the mature catalytically active forms are indicated by an xx for ADAM9, 17 and MMP9. The immature ADAM9 and the mature ADAM10 proteins were not visible. (D) Quantification of experiments in (C). Values shown are mean ± SEM of 6 individual wells and were normalized to vehicle control. ** P <0.01, *** P <0.001, one-way ANOVA with Tukey-Kramer's post-hoc test.

Article Snippet: Following antibodies were purchased: CK2 α (H-286) from Santa Cruz, MMP-9 (G657) and ADAM9 (2099) from Cell Signaling, ADAM17 (T5442) and β-Actin (A5441) from Sigma-Aldrich.

Techniques: Transfection, Expressing, Western Blot

Journal: Biomolecules & Therapeutics

Article Title: 3’-O-Acetyl-24-Epi-7,8-Didehydrocimigenol-3-O-β-D-Xylopryranoside Decreases Amyloid Beta Production in Amyloid Precursor Protein-Transfected HeLa Cells

doi: 10.4062/biomolther.2020.195

Figure Lengend Snippet: Effect of comp 27 on the Expression of ADAM family. (A) Western blot analysis of ADAM9, 10 and 17 after comp 27 treatment. (B) Comp 27 dose-dependently increased the levels of precursor and active ADAM9. (C) Comp 27 dose-dependently increased the levels of precursor and active ADAM10. (D) Comp 27 did not change the levels of both precursor and active ADAM17. * p <0.05, ** p <0.01, and *** p <0.001 compared to the control group.

Article Snippet: Rabbit anti-GAPDH, anti-rabbit horseradish peroxidase linked IgG, anti-ADAM9 antibodies and lysis buffer were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Western Blot

Journal: American Journal of Cancer Research

Article Title: Resveratrol-mediated ADAM9 degradation decreases cancer progression and provides synergistic effects in combination with chemotherapy

doi:

Figure Lengend Snippet: RSV decreases proliferation of ESCC cells. Cell viability was determined by MTT assay in control (shGFP) or ADAM9 knockdown (shADAM9) CE-146T cells (A), KYSE170 (B), Bm7 (C), and A549 (D) with resveratrol treatment for 72 h. Values are means ± SDs for triplicate at each concentration. The IC50 concentrations of RSV in cancer treatment were shown.

Article Snippet: Cell lysates (1.5 mg) were mixed with anti-ADAM9 antibody (4 μl, Cell Signaling, Danvers, MA) for each reaction at 4°C overnight in a rotating wheel, and then 50 μL of protein A/G-coated beads was added and rotated 4°C for 1 hour.

Techniques: MTT Assay, Concentration Assay

Journal: American Journal of Cancer Research

Article Title: Resveratrol-mediated ADAM9 degradation decreases cancer progression and provides synergistic effects in combination with chemotherapy

doi:

Figure Lengend Snippet: RSV promotes ADAM9 protein degradation. (A, B) Western blot analysis of the indicated proteins in lung cancer cells (A) and ESCC cells (B) after RSV treatment for 24 hours. EF1α served as the internal control. (C) RT-qPCR analysis of ADAM9 mRNA expression in cancer cells after RSV treatment for 24 hours. Data represent mean ± SD. (D) Western blot analysis of the indicated proteins in cancer cells after RSV (100 μM) and MG132 (5 μM) treatment. The numbers below the blots indicate the relative protein expression.

Article Snippet: Cell lysates (1.5 mg) were mixed with anti-ADAM9 antibody (4 μl, Cell Signaling, Danvers, MA) for each reaction at 4°C overnight in a rotating wheel, and then 50 μL of protein A/G-coated beads was added and rotated 4°C for 1 hour.

Techniques: Western Blot, Quantitative RT-PCR, Expressing

Journal: American Journal of Cancer Research

Article Title: Resveratrol-mediated ADAM9 degradation decreases cancer progression and provides synergistic effects in combination with chemotherapy

doi:

Figure Lengend Snippet: RSV-induced ADAM9 protein degradation is mediated by the ubiquitin-proteasome pathway. (A, B) HA-Ub was overexpressed in F4 (A) and Bm7 (B) cells treated with RSV (100 μM) and MG132 (5 μM) for 24 hours. Input (left), 10% of the total protein lysate for immunoprecipitation (IP). IP was performed with anti-ADAM9 antibodies, followed by Western blotting with the anti-HA antibody. EF1α, loading control. The numbers below the blots indicate the relative protein expression.

Article Snippet: Cell lysates (1.5 mg) were mixed with anti-ADAM9 antibody (4 μl, Cell Signaling, Danvers, MA) for each reaction at 4°C overnight in a rotating wheel, and then 50 μL of protein A/G-coated beads was added and rotated 4°C for 1 hour.

Techniques: Immunoprecipitation, Western Blot, Expressing

Journal: American Journal of Cancer Research

Article Title: Resveratrol-mediated ADAM9 degradation decreases cancer progression and provides synergistic effects in combination with chemotherapy

doi:

Figure Lengend Snippet: The mechanism of RSV-mediated anticancer effects through promoting ADAM9 protein degradation. Combined RSV with clinical chemotherapeutics can provide synergistic anti-cancer effects.

Article Snippet: Cell lysates (1.5 mg) were mixed with anti-ADAM9 antibody (4 μl, Cell Signaling, Danvers, MA) for each reaction at 4°C overnight in a rotating wheel, and then 50 μL of protein A/G-coated beads was added and rotated 4°C for 1 hour.

Techniques: