Effect of 2.5% O
hypoxia on leptin expression in human coronary artery smooth muscle cells (HCASMCs). (A and B) Leptin protein levels increased, reaching a peak after 4 hours of 2.5% O2 hypoxia. We subjected HCASMCs to hypoxia for 2 to 8 hours. Total cell lysates were harvested and Western blot was performed using anti-leptin antibody. Alpha-tubulin was used as a protein loading control in each lane. *P < 0.01 vs. normoxia control (n = 3). (C) Leptin mRNA level reached a peak after 2 h of 2.5% O2 hypoxia then declined. We used GAPDH as an internal control. *P < 0.01 vs. normoxia control (n = 3).