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. 2014 Mar 25;5:87. doi: 10.3389/fpls.2014.00087

Figure 4.

Figure 4

Subcellular localization analysis of HCC2-mRFP. (A) Five different transgenic lines co-expressing HCC2-mRFP and GFP targeted to mitochondria (mt-GFP) were imaged by laser scanning microscopy, and co-localization of HCC2-mRFP and mt-GFP was found in each line. A representative cell (line 3) is shown, in which co-localization of the green mt-GFP (top panel) and the red HCC2-mRFP signals (middle panel) is depicted as yellow signals after merging the two images (overlay). The overlay also includes a bright-field image of the cell. The image was acquired with a resolution of 1024 × 1024 pixels and a pixel dwell of 3.15 μs. Scale bars correspond to 10 μm each. (B) Mitochondria (M), supernatant (SII) and pellet fractions (PI, PII) were prepared as described in Material and Methods from light-grown WT seedlings or HCC2-mRFP overexpressors (line 5) selected on hyg. The indicated fractions (50 μg each) obtained during the preparations were subjected to 10% SDS-PAGE. Western blot analyses of the same membrane were performed with antibodies directed against mRFP, cytochrome c oxidase subunit 2 (COX2) and the large subunit of Rubisco (RbcL). The total protein in the gel was visualized after the transfer by colloidal Coomassie staining.