Journal: American Journal of Physiology - Cell Physiology

Article Title: Glycolysis and oxidative phosphorylation are essential for purinergic receptor-mediated angiogenic responses in vasa vasorum endothelial cells

doi: 10.1152/ajpcell.00250.2016

Figure Lengend Snippet: Glycolysis and mitochondrial oxidative phosphorylation are necessary for VVEC proliferative and migratory responses. A : proliferation rates were determined in VVEC grown in DMEM/1% FBS in the presence of oligomycin (Oligo; 10 ng/ml), rotenone (Rot; 0.1 μM), 2-deoxyglucose (2-DG; 0.2 mM), and FCCP (0.5 μM) The media with indicated inhibitors were refreshed each second day. Cell proliferation rate was assessed using a fluorescent CyQuant proliferation kit. Values are means ± SE from 3 independent experiments. B : [ 3 H]thymidine incorporation was determined in growth-arrested VVEC (DMEM without serum, 72 h) in response to stimulation with extracellular ATP (100 μM). Cells remained untreated or were treated with 2-DG (2 mM), Rot (0.1 μM), Oligo (100 ng/ml), or FCCP (2 μM) 20 min before stimulation. Values are means ± SE from 3 independent experiments. ## P < 0.01. ### P < 0.001. * P < 0.05, ** P < 0.01, and *** P < 0.001 compared with ATP-treated cells. C : for migration assay, growth-arrested cells were seeded in Boyden chamber-permeable inserts and pretreated with the indicated for 20 min. ATP, MeSADP, or adenosine (Ado; 500 μM) were added to the lower compartment to initiate migration. Number of migrated cells was evaluated as described in methods . Values are means ± SE from 4 independent experiments. ### P < 0.001, control vs. ATP-stimulated cells. * P < 0.05, ** P < 0.01, and *** P < 0.01, untreated vs. treated with inhibitors.

Article Snippet: VVEC were cultured in glass-bottom dishes suitable for fluorescent imaging (MatTek, Ashland, MA) and growth-arrested in serum-free DMEM for 72 h before the experiments.

Techniques: Phospho-proteomics, CyQUANT Assay, Migration, Control