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. 2019 Oct 25;8(11):1319. doi: 10.3390/cells8111319

Figure 1.

Figure 1

Nitric oxide (NO) regulates mitochondria-associated membranes’ (MAMs) integrity in vivo. (A) Representative scheme of strategies used to modulate NO concentration in C57Bl6/JOlash male mice. BH4 (tetrahydrobiopterin; a cofactor of endothelial NO synthase (eNOS), 12.5 mg/kg), l-Name (ω-nitro-l-arginine methyl ester hydrochloride; an inhibitor of eNOS, 25 mg/kg), and a combination of the two were given by intraperitoneal (ip) injection twice the day before and 2 h before euthanasia (n = 5 to 7 mice per condition). Impact of modulating in vivo NO production on (B) liver NO concentration assessed using Daf-FM (15 µM) on fresh homogenates; (C,D) amount of proteins in the MAM fraction isolated from fresh liver using differential ultracentrifugation expressed relative to (C) total and (D) mitochondrial proteins; (E) amount of proteins in the mitochondrial fraction isolated from fresh liver using differential ultracentrifugation expressed relative to total proteins; (F) endoplasmic reticulum (ER)–mitochondria interactions at different distances (from 0 to 50 nm) analyzed from transmission electronic microscopy (TEM) images. (F) Representative TEM images; (G) quantitative analysis of the interactions according to spacing (0–10 nm, 10–20 nm, 20–30 nm, 30–40 nm, 40–50 nm; (H) interactions in a 30–50 nm range; (I) and the number of mitochondria per field (I). A minimum of 10 images (scale bar 0.5 µm) was taken for each mouse (n = 160–260 ER–mitochondria interaction analyzed/mice group condition). £ and #, p < 0.05 vs. control.