Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: β1-Subunit of the calcium-sensitive potassium channel modulates the pulmonary vascular smooth muscle cell response to hypoxia

doi: 10.1152/ajplung.00060.2018

Figure Lengend Snippet: Focal complex expression is greater in pulmonary artery smooth muscle cells (PASMCs) from Kcnmb1−/− compared with Kcnmb1+/+. A: isolated PASMC from both Kcnmb1+/+ and Kcnmb1−/− contract in collagen gel contraction assays. Bars represent means ± SE (n = 3). ***P ≤ 0.001, all samples vs. control (no cells). B: representative images of the expression of focal complexes in isolated peripheral PASMCs 3 h postseeding. Vinculin, red; F-actin, green; nuclei, blue; magnification ×100. Enlarged images, magnification ×200. Quantification of the percentage of cells expressing focal complexes. Bars represent means ± SE (n = 3). ***P ≤ 0.001, 3 h Kcnmb1−/− vs. 3 h Kcnmb1+/+; §§§P ≤ 0.001, 3 h vs. 1 h. C: quantification of the percentage of cells expressing focal complexes in isolated PASMCs left untreated (control), or treated with lithocholic acid (LCA; 45 μM for 30 min), iberiotoxin (IBTX; 55 nM for 1 h), or 5% O2 (hypoxia for 1 h) 3 h postseeding. Bars represent means ± SE (n = 3). ***P ≤ 0.001, Kcnmb1−/− vs. Kcnmb1+/+; §P ≤ 0.05, §§§P ≤ 0.001, treated vs. control.

Article Snippet: Isolated peripheral PASMCs from Kcnmb1 +/+ and Kcnmb1 −/− mice were treated with either an agonist of the β1 subunit of the BK Ca channel, lithocholic acid (LCA; Sigma-Aldrich) ( 7 ) at 45 μM for 30 min, a selective BK Ca channel antagonist, iberiotoxin (Alomone, Jerusalem, Israel) ( 18 ) at 55 nM for 1 h, or hypoxia (5% O 2 ) for 1 h, 3 h postseeding.

Techniques: Expressing, Isolation

Journal: The Journal of Biological Chemistry

Article Title: The Role of ?-Secretase Activating Protein (GSAP) and Imatinib in the Regulation of ?-Secretase Activity and Amyloid-? Generation *

doi: 10.1074/jbc.M112.370924

Figure Lengend Snippet: Effect of imatinib on Aβ generation and Notch processing in cells. a, N2a cells, or b, SHSY5Y-SPA4CT cells were grown in the absence or presence of increasing concentrations of imatinib. 24 h later, medium was removed for Aβ analysis and the cells subject to a cellular viability assay (alamarBlue®). c, membranes from SHSY5Y-SPA4CT cells were incubated for 4 h at 37 °C in the presence of 0.5% DMSO, 10 μm imatinib, or 1 μm GSI and the level of Aβ40 generated was quantified by immunoassay. Data are mean ± S.E. d, SUP-T1 cells were grown in the presence of 0.5% DMSO, 10 μm imatinib, or 1 μm GSI. 24 h later, cells were lysed and subject to Western blot analysis for the Notch-intracellular domain (NICD) or actin. The graph depicts the % change in the level of NICD normalized to the actin control. Data are mean ± S.E. Statistical analysis was conducted using a one-way ANOVA with Dunnett's multiple comparison test (***, p < 0.0001).

Article Snippet: Animals were dosed by intraperitoneal injection with saline (NaCl 0.9%, n = 8) or imatinib mesylate (30 mg/kg, n = 8; Enzo Life Sciences GmbH, Lörrach, Germany) twice daily for 5 days.

Techniques: Cell Viability Assay, Incubation, Generated, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: The Role of ?-Secretase Activating Protein (GSAP) and Imatinib in the Regulation of ?-Secretase Activity and Amyloid-? Generation *

doi: 10.1074/jbc.M112.370924

Figure Lengend Snippet: Effect of imatinib in GSAP siRNA-treated cells. N2a cells were transfected with GSAP siRNA or a scramble siRNA control (Ctrl). 24 h post-transfection medium was replaced with medium containing 0.5% DMSO or 10 μm imatinib. 48 h post-transfection, the effect on GSAP mRNA expression (a) and Aβ generation (b) was determined. Data are mean ± S.E. Statistical analysis was conducted using a one-way ANOVA with Dunnett's multiple comparison test (***, p < 0.0001).

Article Snippet: Animals were dosed by intraperitoneal injection with saline (NaCl 0.9%, n = 8) or imatinib mesylate (30 mg/kg, n = 8; Enzo Life Sciences GmbH, Lörrach, Germany) twice daily for 5 days.

Techniques: Transfection, Expressing

Journal: The Journal of Biological Chemistry

Article Title: The Role of ?-Secretase Activating Protein (GSAP) and Imatinib in the Regulation of ?-Secretase Activity and Amyloid-? Generation *

doi: 10.1074/jbc.M112.370924

Figure Lengend Snippet: Effect of imatinib in vivo. Rats were administered with saline or imatinib (30 mg/kg) twice daily for 5 days or vehicle and a GSI (10 mg/kg) for 4 h. Plasma (a and c) or brain extracts (b and d) were assayed for Aβ40 using a sensitive immunoassay. Statistical analysis was conducted using a one-way ANOVA with Dunnett's multiple comparison test (***, p < 0.0001).

Article Snippet: Animals were dosed by intraperitoneal injection with saline (NaCl 0.9%, n = 8) or imatinib mesylate (30 mg/kg, n = 8; Enzo Life Sciences GmbH, Lörrach, Germany) twice daily for 5 days.

Techniques: In Vivo