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Quantitative analysis of mitochondrial morphologies in human induced pluripotent stem cells for Leigh syndrome

Meshrkey, Fibi, Cabrera Ayuso, Ana, Rao, Raj R., Iyer, Shilpa·2021·Stem Cell Research

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Fig. 1 Mitochondrial morphology of SBG1-(8993 T > G)-hiPSC: a) Representative images of SBG1-(8993 T > G)-hiPSC stained with MTR. From left to right are the phase contrast, Nucblue, RFP, overlay, and skeletonized images including the area of interest of SBG1-(8993 T > G)-hiPSC. Different mitochondrial morphological parameters were determined using MiNA, normalized, and analyzed in comparison to the BJ-hiPSC (control cell line) to quantify: b) the mitochondrial footprint or the skeletal area occupied by the mitochondria c) number of individuals d) number of networks e) mean branch length
Fig. 1 Mitochondrial morphology of SBG1-(8993 T > G)-hiPSC: a) Representative images of SBG1-(8993 T > G)-hiPSC stained with MTR. From left to right are the phase contrast, Nucblue, RFP, overlay, and skeletonized images including the area of interest of SBG1-(8993 T > G)-hiPSC. Different mitochondrial morphological parameters were determined using MiNA, normalized, and analyzed in comparison to the BJ-hiPSC (control cell line) to quantify: b) the mitochondrial footprint or the skeletal area occupied by the mitochondria c) number of individuals d) number of networks e) mean branch lengthCC-BY-NC-ND · From the PDF
Image 2
CC-BY-NC-ND · From the PDF
Fig. 3 Mitochondrial morphology of SBG3-(9185 T > C)-hiPSC: a) Representative images of SBG3-(9185 T > C)-hiPSC stained with MTR. From left to right are the phase contrast, Nucblue, RFP, overlay, and skeletonized images including the area of interest of SBG3-(9185 T > C)-hiPSCs. Different mitochondrial morphological parameters were determined using MiNA, normalized, and analyzed in comparison to the BJ-hiPSC (control cell line) to quantify: b) the mitochondrial footprint or the skeletal area occupied by the mitochondria c) number of individuals d) number of networks e) mean branch length
Fig. 3 Mitochondrial morphology of SBG3-(9185 T > C)-hiPSC: a) Representative images of SBG3-(9185 T > C)-hiPSC stained with MTR. From left to right are the phase contrast, Nucblue, RFP, overlay, and skeletonized images including the area of interest of SBG3-(9185 T > C)-hiPSCs. Different mitochondrial morphological parameters were determined using MiNA, normalized, and analyzed in comparison to the BJ-hiPSC (control cell line) to quantify: b) the mitochondrial footprint or the skeletal area occupied by the mitochondria c) number of individuals d) number of networks e) mean branch lengthCC-BY-NC-ND · From the PDF
Fig. 4 Mitochondrial morphology of SBG4-(10158 T > C)-hiPSCs: a) Representative images of SBG4-(10158 T > C)-hiPSCs stained with MTR. From left to right are the phase contrast, Nucblue, RFP, overlay, and skeletonized images including the area of interest of SBG4-(10158 T > C)-hiPSCs. Different mitochondrial morphological parameters were determined using MiNA, normalized, and analyzed in comparison to the BJ-hiPSC (control cell line) to quantify: b) the mitochondrial footprint or the skeletal area occupied by the mitochondria c) number of individuals d) number of networks e) mean branch length
Fig. 4 Mitochondrial morphology of SBG4-(10158 T > C)-hiPSCs: a) Representative images of SBG4-(10158 T > C)-hiPSCs stained with MTR. From left to right are the phase contrast, Nucblue, RFP, overlay, and skeletonized images including the area of interest of SBG4-(10158 T > C)-hiPSCs. Different mitochondrial morphological parameters were determined using MiNA, normalized, and analyzed in comparison to the BJ-hiPSC (control cell line) to quantify: b) the mitochondrial footprint or the skeletal area occupied by the mitochondria c) number of individuals d) number of networks e) mean branch lengthCC-BY-NC-ND · From the PDF
Image 5
CC-BY-NC-ND · From the PDF
Fig. 6 Mitochondrial membrane potential (MMP) analysis of BJ-hiPSC (control) and five LShiPSC lines. MMP was evaluated by analyzing the intensity of membrane-potential sensitive dye (TMRE). (a) Representative images of BJ and five LS-hiPSCs stained with TMRE. (b) Mean fluorescence intensity (MFI) was calculated based on three images analyzed from three independent dishes from two independent experiments and are shown for (Control BJ-hiPSC in grey; SBG1-(8993 T > G)-hiPSC, SBG2-(8993 T > G)-hiPSC, SBG3-(9185 T > C)-hiPSC in pink; SBG4-(10158 T > C)-hiPSC, and SBG5-(12706 T > C)-hiPSC in blue). *p < 0.05; ****p < 0.0001.
Fig. 6 Mitochondrial membrane potential (MMP) analysis of BJ-hiPSC (control) and five LShiPSC lines. MMP was evaluated by analyzing the intensity of membrane-potential sensitive dye (TMRE). (a) Representative images of BJ and five LS-hiPSCs stained with TMRE. (b) Mean fluorescence intensity (MFI) was calculated based on three images analyzed from three independent dishes from two independent experiments and are shown for (Control BJ-hiPSC in grey; SBG1-(8993 T > G)-hiPSC, SBG2-(8993 T > G)-hiPSC, SBG3-(9185 T > C)-hiPSC in pink; SBG4-(10158 T > C)-hiPSC, and SBG5-(12706 T > C)-hiPSC in blue). *p < 0.05; ****p < 0.0001.CC-BY-NC-ND · From the PDF
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CC-BY-NC-ND · From the PDF

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Authors
Meshrkey, F., Cabrera Ayuso, A., Rao, R. R., & Iyer, S.
Venue
Stem Cell Research
Publisher
Elsevier BV
Published
2021-12
Volume
57
Pages
102572
ISSN
1873-5061
Cited by
24
References
1
Language
en
Funders
  • Arkansas Biosciences Institute
  • Medizinische Universität Innsbruck
  • U.S. Department of Defense (W81XWH-16-1-0181)

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