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. 2008 Feb 11;105(7):2622–2627. doi: 10.1073/pnas.0706790105

Fig. 5.

Fig. 5.

Nuclear PTEN/FOXO pathway regulates HIF-1 transcriptional activity in PTEN-null cancer cells. (A) LnCaP cells infected with adenoviruses. Cell lysates were analyzed by immunoblotting with an anti-PTEN antibody. (B) HIF-1α protein levels in LnCaP cells infected with adenoviruses and subsequently exposed to 21% O2 (N) or 1.5% O2 (H) for 2 h. (C) LnCaP cells were transfected with an HRE-Luciferase reporter gene construct. At 24 h after transfection, cells were infected with adenoviruses and subsequently exposed to 21% O2 or 1.5% O2 for 16 h. Relative luciferase expression is the ratio of luciferase/total protein levels normalized to cells expressing AdNull. (D) LnCaP cells were cotransfected with the HRE-Luciferase reporter gene construct and pLNCX vector control, pLNCX-PTEN (WT), pLNCX-NLS-PTEN, or pLNCX-PTEN (G129R). At 24 h after transfection, cells were exposed to 21% O2 or 1.5% O2 for 16 h. Relative luciferase expression is the ratio of luciferase/total protein levels normalized to pLNCX vector control. (E) LnCaP cells were transfected with an HRE-Luciferase reporter gene construct. At 24 h after transfection, cells were infected with adenoviruses and subsequently exposed to 21% O2 or 1.5% O2 for 16 h. Relative luciferase expression is the ratio of luciferase/total protein levels normalized to cells expressing AdNull. (F) LnCaP cells were transfected with the HRE-Luciferase reporter gene construct and exposed to 21% O2 or 1.5% O2 for 16 h ± 20 nM LMB. Relative luciferase expression is the ratio of luciferase/total protein levels normalized to untreated normoxic cells. (G) U251 cells infected with adenoviruses. Cell lysates were analyzed by immunoblotting with an anti-PTEN antibody. (H) HIF-1α protein levels in U251 cells infected with adenoviruses and subsequently exposed to 21% O2 (N) or 1.5% O2 (H) for 2 h. (I) U251 cells were cotransfected with the HRE-Luciferase reporter gene construct and pLNCX vector control, pLNCX-PTEN (WT), pLNCX-NLS-PTEN, or pLNCX-PTEN (G129R). (J) U251 cells were transfected with the HRE-Luciferase reporter gene construct and exposed to 21% O2 or 1.5% O2 for 16 h ± 20 nM LMB.