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. 2023 Dec 6;24(24):17186. doi: 10.3390/ijms242417186

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© 2023 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Persistency of REC functional mitochondria. (A) Schematic representation of the experiment used to assess the duration of REC mitochondrial function. (B) Intracellular calcium levels were quantified in MELAS neurons (MELAS-N) grown on days 1, 7, 14, and 21 (after receiving REC-donated mitochondria) and in MELAS neurons with high heteroplasmy (n = 9). (C) Representative fluorescence microscopy images of fluo-8 staining in each group of MELAS neurons on days 1, 7, 14, and 21. (D) Representative images of MELAS and REC-treated MELAS neurons stained with JC-1 on days 1, 7, 14, and 21. (E) Quantification of JC-1 fluorescence intensity ratios in REC-treated and untreated MELAS neurons (n = 8). (F) Expression levels of GDF-15 in different groups of neuronal culture media (n = 7). (G) GDF-15 levels were quantified in MELAS neurons grown on days 1, 7, 14, and 21 (after receiving REC-donated mitochondria) and in MELAS neurons with high heteroplasmy (n = 7). (H) Seahorse XFp metabolic flux analysis of mitochondrial ATP production rates and glycolytic ATP production rates in each group of MELAS neurons (n = 8). (I) XFp ATP rate index calculated from data in panel H (n = 8). Data represent the mean ± standard deviation (SD) of three independent experiments. ns, not significant. ** p < 0.01; **** p < 0.0001.

Persistency of REC functional mitochondria. (A) Schematic representation of the experiment used to assess the duration of REC mitochondrial function. (B) Intracellular calcium levels were quantified in MELAS neurons (MELAS-N) grown on days 1, 7, 14, and 21 (after receiving REC-donated mitochondria) and in MELAS neurons with high heteroplasmy (n = 9). (C) Representative fluorescence microscopy images of fluo-8 staining in each group of MELAS neurons on days 1, 7, 14, and 21. (D) Representative images of MELAS and REC-treated MELAS neurons stained with JC-1 on days 1, 7, 14, and 21. (E) Quantification of JC-1 fluorescence intensity ratios in REC-treated and untreated MELAS neurons (n = 8). (F) Expression levels of GDF-15 in different groups of neuronal culture media (n = 7). (G) GDF-15 levels were quantified in MELAS neurons grown on days 1, 7, 14, and 21 (after receiving REC-donated mitochondria) and in MELAS neurons with high heteroplasmy (n = 7). (H) Seahorse XFp metabolic flux analysis of mitochondrial ATP production rates and glycolytic ATP production rates in each group of MELAS neurons (n = 8). (I) XFp ATP rate index calculated from data in panel H (n = 8). Data represent the mean ± standard deviation (SD) of three independent experiments. ns, not significant. ** p < 0.01; **** p < 0.0001.