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Fig. 4 | BMC Biology

Fig. 4

From: MIEF1/2 orchestrate mitochondrial dynamics through direct engagement with both the fission and fusion machineries

Fig. 4

MIEF-mediated mitochondrial fusion is validated by the mito-PAGFP-based mitochondrial fusion assay in WT and Drp1−/− 293T cells. ad The mito-PAGFP-based mitochondrial fusion assay in WT 293T (a, b) and Drp1−/− 293T (c, d) cells. Cells co-transfected with mito-PAGFP (0.5 μg), mito-DsRed (0.2 μg), and either empty vector (0.5 μg), MIEF1 (0.5 μg), or MIEF2 (0.5 μg) were photoactivated in the ROI (white circle, 3 μm diameter) in preactivation images of mitochondria (red). After photoactivation, mito-PAGFP fluorescence (green) intensity and mitochondrial marker (mito-DsRed) were collected at indicated time points (a, c). Mitochondrial fusion was quantified by analyzing changes in fluorescence intensity of photoactivated mito-PAGFP in ROIs at 40 s and 10, 20, and 30 min. The dilution rates (percentage) of the GFP fluorescence intensity at different time points were normalized by the fluorescence intensity at 40 s after photoactivation (b, d) (n represents the number of cells analyzed). ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05

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