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Figure 1 | Molecular Cancer

Figure 1

From: Inhibition of HIF-1alpha activity by homeodomain-interacting protein kinase-2 correlates with sensitization of chemoresistant cells to undergo apoptosis

Figure 1

HIPK2 downregulated HIF-1a expression and protein levels in hypoxia-mimicking condition. (A), RKO-C, RKO-p53i, and T98G cells were transfected with HIPK2-GFP or K221R-GFP expression vectors. Twelve hours after transfection cells were treated with CoCl2 (200 μM) for 24 h. Thirty-six hours after transfection cells were harvested and HIF-1α expression was determined by RT-PCR analysis (Total time treatments: 36 h transfection, 24 h CoCl2). Aldolase (aldo) was used as loading control. One representative experiment from three independent experiments is shown. (B), Nuclear extracts from cells treated as in (A) were analyzed by Western immunoblotting with anti-HIF-1α antibody. Anti-Hsp70 was used as protein loading control. (C), RKO-C cells were transfected with HIPK2-GFP or GFP-empty vectors. Twelve hours after transfection cells were treated with CoCl2 (200 μM) for 20 h before adding cycloheximide (CHX) (40 μg/ml, t = 0). Nuclear extracts were analyzed by Western immunoblotting 60, 120, and 180 minutes following CHX treatment, with anti-HIF-1α antibody. Anti-Hsp70 was used as protein loading control. (D), The relative amount of HIF-1α protein levels from panel (C) was quantitated by densitometric analysis and plotted. The half-life of HIF-1α is shown by dotted lines.

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Molecular Cancer

ISSN: 1476-4598