Figure 5 | Scientific Reports

Figure 5

From: Peptide-mediated delivery of donor mitochondria improves mitochondrial function and cell viability in human cybrid cells with the MELAS A3243G mutation

Figure 5

Mitochondrial morphology, mitochondria-shaping proteins and mitochondrial biogenesis in rescued cells after 4 days of treatment. (A) Morphology of mitochondria stained with MitoTracker Green observed by confocal microscopy. (B) Quantification of tubular and fragmented mitochondria was further analyzed using a semi-automatic system. Images from three independent areas containing approximately 200–300 mitochondria from about 6–8 cells each were analyzed from each group (n = 3). (C) Western blot analysis of mitochondrial fusion proteins OPA1 (including long and short OPA1 isoforms) and MFN2, fission protein DRP1 and mitochondrial marker proteins Tom20 and Tim23. (D) Protein expression was quantified by densitometry and normalized to GAPDH (n = 3). (E) Mitochondrial amount analyzed by NAO staining and flow cytometry. (F) Expression of mitochondrial biogenetic genes PGC-1α, NRF-1 and Tfam analyzed by RT-PCR. Relative expression levels were determined relative to β-actin. *p < 0.05, compared to control group; + p < 0.05, compared to MELAS group; # p < 0.05, compared to Pep-1 group; control cybrid cells (Con); MELAS cybrid cells (MELAS); MELAS cybrid cells treated with Pep-1 alone (Pep-1) or Pep-1-labeled mitochondria (Pep-1-Mito).

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