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. 2004 Sep;24(17):7469–7482. doi: 10.1128/MCB.24.17.7469-7482.2004

FIG. 4.

FIG. 4.

ATF4 is translationally induced after hypoxic stress. (A) Immunoblot analysis of ATF4 and HIF-1α protein content in HeLa cells exposed to hypoxia or left untreated for the indicated period of time. Actin serves as a loading control. (B) Immunoblot of ATF4 and BiP in HeLa cells treated with hypoxia for the indicated period of time in the presence or absence of the transcriptional inhibitor ActD (100 μM) added 5 min before treatment. Similar results were obtained by using another transcriptional inhibitor, DRB at 100 μM (data not shown). Actin serves as a loading control. (C) Immunoblot of ATF4 and HIF-1α in HeLa cells treated with hypoxia for the indicated period of time in the presence or absence of the translational inhibitor CHX (100 μM). Actin serves as a loading control. (D) Immunoblot of ATF4 and HIF-1α in HeLa cells exposed to hypoxia for the indicated period of time. Cells were first treated with hypoxia, followed by the addition of the translational inhibitor CHX for 15 min, 30 min, or 1 h. Actin served as a loading control.