FIG. 8.

The growth rate of wild-type cells treated with glycolytic inhibitors parallels the growth rate observed in HIF-1α-null cells. Following seeding at moderate density (2 × 10⁵) and plating overnight, cells were washed once with PBS and then cultured in DMEM–high-glucose medium supplemented with 25 mM HEPES (pH 7.4) containing either no drug (NO), 6 mM oxamate (OX), or 6 mM 2-deoxyglucose (2-DEOXY). After 24 h of incubation at hypoxia, triplicate plates of cells per treatment were harvested by trypsinization and counted by using a hemacytometer. This graph is representative of the changes in growth rates ± the SEM observed in three independent experiments. Treatment of wild-type cells with either oxamate or 2-deoxyglucose reduces the growth of wild-type cells to levels similar to those observed in cells deleted for HIF-1α, indicating that the proper regulation of glycolysis is required for maximal cell growth during hypoxia.