Hypoxia-inducible factor-1 alpha as a therapeutic target for primary effusion lymphoma
Fig 2
Effect of HIF-1α knockdown on glycolysis and lipid biogenesis.
(A) Levels of extracellular lactate produced after 24 hours in normoxia (N) or hypoxia (H), expressed as fold change relative to cells containing shScr under normoxia. (B) Protein levels of HIF-1α regulated glycolytic genes after 48 hours in normoxia or hypoxia as assessed by Western blot. β-actin is shown as a loading control. (C) mRNA levels of HIF-1α regulated glycolytic genes measured by RT-qPCR after 48 hours in N or H. mRNA levels are normalized to that of 18S ribosomal RNA and are expressed as fold change relative to cells containing shScr under N. (D) Levels of miR210 measured by taqman RT-qPCR assay. RNA levels are normalized to that of internal control RNU43 and expressed as fold change relative to shScr cells in normoxia. (E) Levels of phospholipids produced in shHIF-1 cells relative to shScr cells in normoxia. (F) mRNA levels of SIAH2 in BCBL-1 cells normalized to 18S and expressed as fold change relative to shScr cells under N. (G) Protein levels of OGDH2 after 48 hours in N or H. Numbers below the blot for OGDH2 represent protein levels normalized to β-actin and expressed as fold change relative to shScr cells in N. Western blots were done on lysates from three independent experiments and representative blots are shown. Error bars represent standard deviations from at least 3 independent experiments. Statistically significant differences between shScr and shHIF-1 cells are indicated. *P ≤0.05, **P ≤ 0.01 (2-tailed t-test, paired).