American Society of Hematology

Figure 2.

$Figure 2. Amphotericin B inactivates HIF-1 without changing protein levels of HIF-1 subunits. (A-D) AmB-induced HIF-1 inhibition. Luciferase reporter plasmids containing wild EpoE or mutated EpoE were transfected into Hep3B and HEK293 cells. After incubation under normoxic or hypoxic conditions for 16 hours with sodium deoxycholate or AmB, luciferase activities were measured. Results are relative values versus the normoxic control and are plotted as the means ± SD of 12 experiments. *P < .01 versus the hypoxic control. (E) No changes in expression and nuclear translocation of HIF-1 subunits. Hep3B and HEK293 cells were incubated under normoxic or hypoxic conditions for 16 hours with sodium deoxycholate (10 μg/mL) or AmB (1.25 to 10 μg/mL). HIF-1α and ARNT proteins in total cell lysates (left panel) or in nuclear fractions (right panel) were analyzed by Western blotting. β-actin or NF-κB p50 protein was used as a loading control for total cell lysates or nuclear fractions, respectively. The data shown are representative of 3 separate experiments.$

Amphotericin B inactivates HIF-1 without changing protein levels of HIF-1 subunits. (A-D) AmB-induced HIF-1 inhibition. Luciferase reporter plasmids containing wild EpoE or mutated EpoE were transfected into Hep3B and HEK293 cells. After incubation under normoxic or hypoxic conditions for 16 hours with sodium deoxycholate or AmB, luciferase activities were measured. Results are relative values versus the normoxic control and are plotted as the means ± SD of 12 experiments. ^*P < .01 versus the hypoxic control. (E) No changes in expression and nuclear translocation of HIF-1 subunits. Hep3B and HEK293 cells were incubated under normoxic or hypoxic conditions for 16 hours with sodium deoxycholate (10 μg/mL) or AmB (1.25 to 10 μg/mL). HIF-1α and ARNT proteins in total cell lysates (left panel) or in nuclear fractions (right panel) were analyzed by Western blotting. β-actin or NF-κB p50 protein was used as a loading control for total cell lysates or nuclear fractions, respectively. The data shown are representative of 3 separate experiments.

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