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. 2014 Mar 24;111(14):5331–5336. doi: 10.1073/pnas.1317242111

Fig. 2.

Fig. 2.

Reciprocal relationship between DNM and HIF-1α. (A) Inhibition of Dynamin rapidly stabilizes HIF-1α. Representative Western blot of HIF-1α is shown. (B) Dynamin inhibition increases ODDD-HIF-1α-luc fusion protein. (C) Dominant-negative DNM (K44A-DNM2) rescued DNM-mediated down-regulation of HIF-1α in hypoxia. Western blot for HIF-1α is shown. Normoxia (N; lanes 1 and 2), hypoxia (H; lanes 3 and 4). (D) DNM2 overexpression inhibits HIF-1α. (E) DNM2 knockdown increases HIF-1α and HIF-2 α levels (representative Western blot). (F) Iron levels were indirectly measured by quenching of calcein-AM fluorescence. (G) Stabilization of HIF-1α by Dynamin inhibition is reversed by FAC treatment (representative Western blot). (H) Dynamin inhibition blocks polyubiquitination of HIF-1α. Proteasome inhibitor, MG132, was used to block degradation of HIF-1α. Dyna, Dynasore; Sc, scrambled control.