Extended Data Fig. 10: Measurement of mitochondrial fusion status.
From: Dual regulation of mitochondrial fusion by Parkin–PINK1 and OMA1
(a) Western blotting of Drp1 KO MEFs transduced with lentiviruses carrying Opa1 and/or Mfn1. The asterisk indicates non-specific bands of anti-Mfn1 antibodies. Quantification of band intensity is shown (mean ± SD, n = 3). (b) Drp1 KO MEFs carrying matrix-targeted photoactivatable GFP (mito-PA-GFP), along with Opa1 and/or Mfn1, were stained with tetramethylrhodamine ethyl ester (TMRE). mito-PA-GFP in a single mitochondrion (indicated by the boxes) in the cell periphery was photoactivated, and images were obtained every 1 min at a single focal plane. Photoactivation was performed every 1 min on the same mitochondrion to maintain signal intensity. Representative images before and after (0 and 15 min) photoactivation are shown. (c) The mitochondrial fusion status was calculated based on the relative area containing photoactivated mito-PA-GFP signals over the total mitochondria stained with TMRE at 15 min (mean ± SD, n = 35 cells for Drp1 KO, 32 cells for Drp1 KO + Opa1, 34 cells for Drp1 KO + Mfn1, and 31 cells for Drp1 KO + Opa1 + Mfn1). Statistical analysis was performed using one-way ANOVA with post-hoc Tukey test: *p < 0.05, ***p < 0.001.
