Fig. 2

Interference with O-GlcNAc metabolic enzymes modulated mitochondrial metabolic function and affects mitophagy in hyperoxia-exposed RLE-6TN cells. a CCK-8 assay of cell proliferation after exposing RLE-6TN cells to hyperoxia and treatment with 1 μmol L^–1 Thiamet G. b CCK-8 assay of cell proliferation after exposing RLE-6TN cells to hyperoxia and treatment with 1 μmol L^–1 OSMI-1. c CCK-8 assay of cell proliferation after exposing RLE-6TN cells to hyperoxia and treatment with 20 μmol L^–1 UDP-GlcNAc. d–f Metabolic profiles determined using extracellular flux analysis (Seahorse XF). The oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) in each condition are shown, following the addition of oligomycin (0.5 μM), FCCP (0.25 μM), and antimycin (1 μM) and rotenone (1 μM), glutamine (2 mM), 10 mM glucose, 1 μM oligomycin, and 50 mM 2-DG according to the protocol. g Graphs of JC-1 fluorescence as a measure of the cell membrane potential. Red indicates a normal membrane potential, whereas green indicates a reduced membrane potential (magnification × 400; scale bar = 50 µm). Data are shown as the mean ± SEM (n = 6) and were analyzed using two-way ANOVA followed by Tukey’s post-hoc tests. *P < 0.05, **P < 0.01. NO: normal air-exposed group; HO: hyperoxia-exposed group. HO + TG: 48 h hyperoxia-exposed and Thiamet G-treated group; HO + OS: 48 h hyperoxia-exposed and OSMI-1-treated group; UDP: 48 h hyperoxia-exposed and UDP-GlcNAc-treated group; FCCP: carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone