Figure 2.
Figure 2. Increased oxidative stress and disturbed mitochondrial morphology in NLRX1 KO mice during IRI. Digital image analysis of immunohistochemistry for (A) 4-hydroxynonenal (4-HNE) and (B) nitrosylated tyrosine on kidney tissue sections from WT and KO animals (white and black bars, respectively, n = 8 per group) at the indicated time points during IRI (left). Representative images are presented right for each stain. Bars, 100 µm. All data are expressed as mean ± SEM and were analyzed using a Mann-Whitney U test (*, P < 0.05; **, P < 0.01). contr, contralateral kidney. (C–F) Transmission electron microscopic imaging of kidney sections obtained from (C and E) WT and (D and F) NLRX1 KO animals. Proximal tubules in contralateral kidneys from (C) WT and (D) NLRX1 KO animals show normal mitochondrial morphology. (E) Kidney section from a WT animal at day 1 after ischemia shows mild mitochondrial fragmentation (indicated by black arrows). (F) Kidney section from an NLRX1 KO animal at day 1 after ischemia shows extensive mitochondrial fragmentation (indicated by black arrow) and mitochondrial collapse (indicated by white arrows). Bars, 2 µm. (C′–F′) High-magnification imaging of mitochondria from (C′ and D′) contralateral kidneys and (E′ and F′) injured kidneys. (G) Semiquantitative scoring of mitochondrial damage in WT and NLRX1 KO mice at day 1 during IRI (white and black bars, respectively, n = 3 per group). Examples of each score are presented. Data are expressed as mean ± SEM and were analyzed using a Mann-Whitney U test (*, P < 0.05). White arrows indicate sites of membrane rupture. (H) Transmission electron microscopic imaging of a kidney section obtained from an NLRX1 KO animal at day 1 after ischemia showing a phagosome-like structure containing mitochondria. Mitochondrial fragmentation (black arrows) and mitochondrial collapse (white arrows) are indicated; n, nucleus. (I) Transcription of glutathione peroxidase (GPX) 1 and 3, peroxiredoxin 1 (PRDX1), and superoxide dismutase 2 (SOD2) in kidney tissue from WT and KO animals (white and black bars, respectively, n = 8 per group) at the indicated time points during IRI. Bars, 500 nm, 1 µm, or 2 µm as indicated. Data are expressed as mean ± SEM and were analyzed using a Mann-Whitney U test (*, P < 0.05).

Increased oxidative stress and disturbed mitochondrial morphology in NLRX1 KO mice during IRI. Digital image analysis of immunohistochemistry for (A) 4-hydroxynonenal (4-HNE) and (B) nitrosylated tyrosine on kidney tissue sections from WT and KO animals (white and black bars, respectively, n = 8 per group) at the indicated time points during IRI (left). Representative images are presented right for each stain. Bars, 100 µm. All data are expressed as mean ± SEM and were analyzed using a Mann-Whitney U test (*, P < 0.05; **, P < 0.01). contr, contralateral kidney. (C–F) Transmission electron microscopic imaging of kidney sections obtained from (C and E) WT and (D and F) NLRX1 KO animals. Proximal tubules in contralateral kidneys from (C) WT and (D) NLRX1 KO animals show normal mitochondrial morphology. (E) Kidney section from a WT animal at day 1 after ischemia shows mild mitochondrial fragmentation (indicated by black arrows). (F) Kidney section from an NLRX1 KO animal at day 1 after ischemia shows extensive mitochondrial fragmentation (indicated by black arrow) and mitochondrial collapse (indicated by white arrows). Bars, 2 µm. (C′–F′) High-magnification imaging of mitochondria from (C′ and D′) contralateral kidneys and (E′ and F′) injured kidneys. (G) Semiquantitative scoring of mitochondrial damage in WT and NLRX1 KO mice at day 1 during IRI (white and black bars, respectively, n = 3 per group). Examples of each score are presented. Data are expressed as mean ± SEM and were analyzed using a Mann-Whitney U test (*, P < 0.05). White arrows indicate sites of membrane rupture. (H) Transmission electron microscopic imaging of a kidney section obtained from an NLRX1 KO animal at day 1 after ischemia showing a phagosome-like structure containing mitochondria. Mitochondrial fragmentation (black arrows) and mitochondrial collapse (white arrows) are indicated; n, nucleus. (I) Transcription of glutathione peroxidase (GPX) 1 and 3, peroxiredoxin 1 (PRDX1), and superoxide dismutase 2 (SOD2) in kidney tissue from WT and KO animals (white and black bars, respectively, n = 8 per group) at the indicated time points during IRI. Bars, 500 nm, 1 µm, or 2 µm as indicated. Data are expressed as mean ± SEM and were analyzed using a Mann-Whitney U test (*, P < 0.05).

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