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. 2015 Jan 20;112(5):1553–1558. doi: 10.1073/pnas.1419767112

Fig. 1.

Fig. 1.

Hypoxia-inducible Higd1a directly binds to highly purified cytochrome c oxidase (hpCcO). (A) Heat map of three genes (Upper) identified as relatively rapid and transiently induced in response to hypoxia in rat neonatal cardiomyocytes, compared with genes known to be hypoxia inducible (Pfkl, Hk2, Hmox1, and Vegfa) (Lower). (B) Expression of the Higd1a protein was elevated in response to hypoxia. (C) In vitro pull-down assay with amylose resin revealed direct binding between MBP-Higd1a and the hpCcO from bovine heart. Loading controls for the hpCcO and MBP-fusion proteins are shown in immunoblots for anti-CcO subunits and CBB staining, respectively. (D) MBP-Higd1a directly integrates into hpCcO. Mixed MBP-fusion proteins and hpCcO containing 0.2% n-decyl-β-d-maltoside (DM) were resolved by blue native PAGE (BN-PAGE), followed by immunoblotting with anti-Cox4 to detect CcO and anti-Higd1a to detect Higd1a.