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. 2019 Nov 25;10:1449. doi: 10.3389/fpls.2019.01449

Figure 8.

Figure 8

MITs are important for mitochondrial iron acquisition/import. (A) Separation of 20 µg solubilized mitochondrial fraction of Col-0 and the mit1 −− /mit2 + − mutant grown in Fe-sufficient (+Fe) or Fe drop-out media (-Fe) by blue-native gel electrophoresis (BN-PAGE). (B) In-gel enzyme activity assay for complex I on mitochondrial extract separated by BN-PAGE. The arrows indicate complex I and its supercomplexes. Non-specific staining at the bottom of the gel was used as the loading control. The staining in the WT and the mutant was quantified using ImageJ software. (C) Immunoblots showing aconitase protein levels in mitochondrial and total protein extracts. Coomassie stained internal proteins were used as a loading control. Total mitochondrial (D) Fe content, (E) Zn content, and (F) Mn content as measured by ICP-MS (shown is the mean of three technical replicates; error bars represent standard deviation). Significance was assessed using the student’s t-test. An asterisk represents a p value < 0.05. The mitochondria for all the experiments (B-F) were isolated from 2.5-week-old seedlings grown on Fe drop-out media.