Figure 4.

Prolonged hypoxia increases glutamine metabolism in PDAC cells, and HIF‐2α can promote non‐canonical glutamine metabolism in chronic hypoxic conditions. (A) Time course of glutamine consumption at 1%, 3% or 20% O₂, each time data point is an average of triplicate experiments. (B) Panc‐1 and Capan‐2 were incubated for 48 hrs at 1%, 3% or 20% O₂, GLS1, GOT1 and GOT2 protein were measured by Western blot. β‐Actin was used as loading control. (C) Panc‐1 and Capan‐2 were incubated for 48 hrs at 1%, 3% or 20% O₂, GLS1, GOT1 and GOT2 mRNA were measured by qRT‐PCR. Data are presented as mean ± S.D. from three independent experiments. (D and F) Si‐HIF‐2α‐transfected Panc‐1 and Capan‐2 cultured at 1% or 3%O₂ for 48 hrs. The level of HIF‐2α and glutamine metabolism enzymes mRNA and protein were determined by qRT‐PCR (mRNA) and Western blot (protein), and β‐actin was used as loading control. Data are presented as mean ± S.D. from three independent experiments. *P < 0.05, **P < 0.01.