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

Article Title: Hypoxia induces arginase II expression and increases viable human pulmonary artery smooth muscle cell numbers via AMPKα 1 signaling

doi: 10.1152/ajplung.00117.2016

Figure Lengend Snippet: Pharmacologic inhibition of AMPK prevents hypoxia-induced arginase II protein expression, whereas AMPK activation increases arginase II protein expression. Human PASMCs were treated with the AMPK inhibitor Compound C, the cell-permeable AMPK activator AICAR, or vehicle (DMSO for the AMPK inhibitors or dH2O for AMPK activator, N1-(β-d-ribofuranosyl)-5-aminoimidazole-4-carboxamide (AICAR), and incubated in 21% O2 (normoxia) or 1% O2 (hypoxia) for 48 h. Protein was assayed for arginase II protein expression by Western blot analysis and was quantified by densitometry. A: representative Western blots are shown for arginase II and β-actin. B: data are shown as a fold change ± SE relative to vehicle-treated, normoxia-exposed human PASMC controls (n = 4). *Significant difference compared with vehicle-treated normoxia, P < 0.05. †Significant difference compared with vehicle-treated hypoxia, P < 0.0001.

Article Snippet: In some experiments, AMPK inhibitors 6-[4-(2-piperidin-1-yl-ethoxy)-phenyl)]-3-pyridin-4-yl-pyrrazolo[1,5-a]-pyrimidine (Compound C, 10 μM, cat. no. BML-EI369, lot no. 12171240, Enzo Life Sciences, Farmingdale, NY) or adenine 9-β- d -arabinofuranoside (Ara-A, 100 μM, cat. no. A5762, lot no. BCBL 6590V; Sigma Gel, Jandel Scientific, San Rafael, CA), the AMPK activator, N 1 -(β- d -ribofuranosyl)-5-aminoimidazole-4-carboxamide (AICAR; 150 μM, cat. no. sc-200659, lot no. A2413; Santa Cruz Biotechnology, Santa Cruz, CA), or vehicle (DMSO for the AMPK inhibitors or dH 2 O for AICAR) was added to the medium.

Techniques: Inhibition, Expressing, Activation Assay, Incubation, Western Blot