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. 2017 Nov 28;37(24):e00337-17. doi: 10.1128/MCB.00337-17

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Copyright © 2017 Ishimoto et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 6 — Mitochondrial abnormalities in human ADPKD cyst-derived cells with a heterozygous PKD1 mutation. (A) PKD1 expression in cyst-derived cells containing a heterozygous PKD1 mutation (WT 9-7) compared to a normal tubular cell line (RCTEC-LTA) (each group, n = 3). Results represent the relative ratio. (B) mtDNA copy number in WT 9-7 relative to RCTEC-LTA (each group, n = 3). Results represent the relative ratio. (C) Western blot analysis of PGC-1α levels of WT 9-7 and RCTEC-LTA (each group, n = 3). The bar graph shows the relative ratio of protein expression calibrated by histone H1 in normal and cyst-derived cells. (D) mRNA expression of PGC-1α in WT 9-7 and RCTEC-LTA (each group, n = 3). The bar graph shows the relative ratio of mRNA expression calibrated by GAPDH in normal and cyst-derived cells. (E) MitoTracker Green staining of RCTEC-LTA and WT 9-7. Mitochondrial elongation was evaluated using ImageJ software, and units represent the major axis to minor axis ratio (each group, n = 25). (F) Evaluation of mitochondrial membrane potential. The bar graph shows the percentage of depolarized cells, showing depolarization of inner mitochondrial membrane potential in WT 9-7 compared with that in RCTEC-LTA (each group, n = 3). (G) Evaluation of intracellular ROS (superoxide). The x and y axes represent the cell number and the ROS signaling strength, respectively, with ROS-negative cells (M1) and ROS-positive cells (M2) identified. The ROS-positive cell number in WT 9-7 is compared with that in RCTEC-LTA. The bar graph shows the percentage of ROS-positive cells (each group, n = 3). (H) Evaluation of mitochondrial superoxide by MitoSOX Red staining. The box plot shows the signal intensity. Mitochondrial superoxide levels in WT 9-7 are compared to those in RCTEC-LTA (each group, n = 25). Results represent the means ± standard deviations. **, P < 0.01; ***, P < 0.001.

FIG 6 — Mitochondrial abnormalities in human ADPKD cyst-derived cells with a heterozygous PKD1 mutation. (A) PKD1 expression in cyst-derived cells containing a heterozygous PKD1 mutation (WT 9-7) compared to a normal tubular cell line (RCTEC-LTA) (each group, n = 3). Results represent the relative ratio. (B) mtDNA copy number in WT 9-7 relative to RCTEC-LTA (each group, n = 3). Results represent the relative ratio. (C) Western blot analysis of PGC-1α levels of WT 9-7 and RCTEC-LTA (each group, n = 3). The bar graph shows the relative ratio of protein expression calibrated by histone H1 in normal and cyst-derived cells. (D) mRNA expression of PGC-1α in WT 9-7 and RCTEC-LTA (each group, n = 3). The bar graph shows the relative ratio of mRNA expression calibrated by GAPDH in normal and cyst-derived cells. (E) MitoTracker Green staining of RCTEC-LTA and WT 9-7. Mitochondrial elongation was evaluated using ImageJ software, and units represent the major axis to minor axis ratio (each group, n = 25). (F) Evaluation of mitochondrial membrane potential. The bar graph shows the percentage of depolarized cells, showing depolarization of inner mitochondrial membrane potential in WT 9-7 compared with that in RCTEC-LTA (each group, n = 3). (G) Evaluation of intracellular ROS (superoxide). The x and y axes represent the cell number and the ROS signaling strength, respectively, with ROS-negative cells (M1) and ROS-positive cells (M2) identified. The ROS-positive cell number in WT 9-7 is compared with that in RCTEC-LTA. The bar graph shows the percentage of ROS-positive cells (each group, n = 3). (H) Evaluation of mitochondrial superoxide by MitoSOX Red staining. The box plot shows the signal intensity. Mitochondrial superoxide levels in WT 9-7 are compared to those in RCTEC-LTA (each group, n = 25). Results represent the means ± standard deviations. **, P < 0.01; ***, P < 0.001.