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. 2016 Dec 6;173(2):956–969. doi: 10.1104/pp.16.01641

Figure 2.

Figure 2.

AtSLP2 and AtMIA40 are located in the mitochondrial IMS. A, Stably transfected Arabidopsis roots expressing 35Spro::AtSLP2-GFP (green) are stained with MitoTracker Deep Red FM (purple) to illuminate mitochondria. Representative individual, merged, and bright field images are shown. B, Intact mitochondria and peroxisomes were isolated from dark-grown wild-type Arabidopsis cell culture. Immunoblotting of the purified mitochondrial (Mt) and peroxisomal (Px) fractions were performed using 1:100 antimitochondrial pyruvate dehydrogenase complex E1α (anti-E1α; mitochondrial matrix) and 1:200 anticatalase (peroxisome) IgG. Each lane contains 15 µg of total protein. C, Isolated mitochondria (Mt) were further subfractionated into OMM and IMS and immunoblotted using 1:5000 anti-H protein of Gly decarboxylase complex (anti-GDC-H; mitochondria matrix marker), 1:5,000 antivoltage-dependent anion-selective channel protein 1 (anti-VDAC1; OMM marker) and 2 µg/mL anticytochrome c (anticyt c; IMS marker) IgG. Each lane contains 10 µg of total protein. Immunoblotting was performed in conjunction with affinity-purified anti-AtSLP2 IgG (Uhrig and Moorhead, 2011) and anti-AtMIA40 crude immune serum (Supplemental Fig. S4) used at 1.2 µg/mL and a 1:1,000 dilution, respectively.