Figure 5.
Figure 5. Loss of NLRX1 disrupts mitochondrial morphology but increases mitochondrial activity. (A) Confocal microscopic images of cEmpty (top) and cNLRX1 cells (bottom) labeled with MitoTracker Deep Red (red) at the indicated time points. Nuclei (blue) were counterstained; bars, 10 µm. White arrows indicate examples of mitochondrial fragmentation. Conditions were examined in triplicate; representative images are shown. reox, reoxygenation. (B) Analysis of mitochondrial membrane potential using JC-1 labeling under normoxic conditions or at indicated time points after hypoxia for cEmpty cells (open circles) and cNLRX1 cells (closed circles). All conditions were examined in sixfold. Representative data from three independent experiments are shown. (C) Seahorse Bioanalyzer OCR readout of a typical mitochondrial function assay using cEmpty (open circles) and cNLRX1 (closed circles) without adjustment for cell number. AA, antimycin A; rot, rotenone. Conditions were determined in sixfold; representative data from three independent experiments are shown. (D) OCR data from C adjusted for cell input for cEmpty cells (white bars) and cNLRX1 cells (black bars). (E) Seahorse Bioanalyzer extracellular acidification rate (ECAR) readout of a typical mitochondrial function assay using cEmpty cells (open circles) and cNLRX1 cells (closed circles) without adjustment for cell number. Conditions were determined in sixfold. Representative data from three independent experiments are shown. (F) Complex II/III activity determined in isolated kidney mitochondria from WT (white bars) and NLRX1 KO mice (black bars; n = 3 per group). (G) Enzyme activity of citrate synthase, hexokinase, LDH, and SCHAD determined in normoxic cEmpty and cNLRX1 cells. All conditions were examined in triplicate; representative data from three independent experiments are shown. (H) ATP levels in cEmpty cells (white bars) and cNLRX1 cells (black bars) were measured at the indicated time points and adjusted to total protein contents per sample. Conditions were examined in sixfold. Representative data from three independent experiments are shown. All data are expressed as mean ± SEM and were analyzed using an unpaired t test except enzyme activity by Mann-Whitney U test (*, P < 0.05; **, P < 0.01; ***, P < 0.001).

Loss of NLRX1 disrupts mitochondrial morphology but increases mitochondrial activity. (A) Confocal microscopic images of cEmpty (top) and cNLRX1 cells (bottom) labeled with MitoTracker Deep Red (red) at the indicated time points. Nuclei (blue) were counterstained; bars, 10 µm. White arrows indicate examples of mitochondrial fragmentation. Conditions were examined in triplicate; representative images are shown. reox, reoxygenation. (B) Analysis of mitochondrial membrane potential using JC-1 labeling under normoxic conditions or at indicated time points after hypoxia for cEmpty cells (open circles) and cNLRX1 cells (closed circles). All conditions were examined in sixfold. Representative data from three independent experiments are shown. (C) Seahorse Bioanalyzer OCR readout of a typical mitochondrial function assay using cEmpty (open circles) and cNLRX1 (closed circles) without adjustment for cell number. AA, antimycin A; rot, rotenone. Conditions were determined in sixfold; representative data from three independent experiments are shown. (D) OCR data from C adjusted for cell input for cEmpty cells (white bars) and cNLRX1 cells (black bars). (E) Seahorse Bioanalyzer extracellular acidification rate (ECAR) readout of a typical mitochondrial function assay using cEmpty cells (open circles) and cNLRX1 cells (closed circles) without adjustment for cell number. Conditions were determined in sixfold. Representative data from three independent experiments are shown. (F) Complex II/III activity determined in isolated kidney mitochondria from WT (white bars) and NLRX1 KO mice (black bars; n = 3 per group). (G) Enzyme activity of citrate synthase, hexokinase, LDH, and SCHAD determined in normoxic cEmpty and cNLRX1 cells. All conditions were examined in triplicate; representative data from three independent experiments are shown. (H) ATP levels in cEmpty cells (white bars) and cNLRX1 cells (black bars) were measured at the indicated time points and adjusted to total protein contents per sample. Conditions were examined in sixfold. Representative data from three independent experiments are shown. All data are expressed as mean ± SEM and were analyzed using an unpaired t test except enzyme activity by Mann-Whitney U test (*, P < 0.05; **, P < 0.01; ***, P < 0.001).

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal