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. 2013 Oct 21;110(45):18168–18173. doi: 10.1073/pnas.1311382110

Fig. 1.

Fig. 1.

Gam1 WT decreases cellular levels of VHL, stabilizes HIF 1α, and increases the transactivational function of HIF 1α. (A) Immunoblotting (IB) showing Glut1 and HIF1α stabilization. HeLa Tet-on cells were induced (DOX) to express Myc Gam1 WT. Lysates were made in SDS lysis buffer at the indicated times and probed as indicated. Loading controls: Vinculin and Tubulin. (B) Luciferase assay. HeLa cells transfected with HRE (Hypoxia Responsive Element)-Luciferase reporter plasmid together with either empty vector or myc Gam1 wild-type (WT). Results are reported as fold induction of luciferase activity in relative light units per second (RLU/s) normalized to the control (empty vector) where n = 3 ± SD. (C) Quantitative RT-PCR. Results are reported as fold induction of CAIX mRNA relative to the control (empty vector) and normalized respectively to GADPH mRNA (to which the arbitrary value of 1 was assigned) where n = 3 ± SD. (D) IB of VHL and HIF 1α in HeLa cells transiently transfected with Myc Gam1 WT and Myc Gam1 LL/AA. Loading control: Tubulin and Vinculin.