Figure 6. BDH2 is Required for Mitochondrial Iron Homeostasis.

(A–F) FL5.12 cells expressing a Bdh2 or control shRNA were analyzed for mitochondrial iron levels by colorimetric analysis (A), for mitochondrial iron uptake (B), for RPA (C) and RPAC (D) fluorescence by fluorometry, for DHR 123 oxidation in the presence or absence of H₂O₂ by flow cytometry (E), and for activities of mitochondrial aconitase, succinate dehydrogenase and citrate synthase (F).

(G) In vitro mitochondrial iron import. (Left) Proteolytically treated cytosolic extracts from control or Bdh2 shRNA-treated FL5.12 cells were added to purified mitochondria, and iron import was determined by monitoring mitochondrial incorporation of ⁵⁵FeCl₃. (Right) Purified mitochondria were treated with 2,3-DHBA, 2,5-DHBA, enterobactin or DFO, and iron import was monitored. See also Figures S3F-J and S5.