Fig. 3.

HINT2 overexpression alleviated OGD/R injury-induced mitochondrial dysfunction by restraining mitochondrial fragmentation. CMECs were isolated from 6- to 8-week-old WT mice, transfected with ADV-NC or ADV-HINT2, and subjected to OGD/R injury. Mdivi-1 (5 μM) was added to the ADV-NC group during OGD/R injury to inhibit mitochondrial fission. FCCP (1 μM) was applied to ADV-HINT2 cells during OGD/R injury for 2 h to reintroduce mitochondrial fission. Mitochondrial membrane potential was measured using JC-1 fluorescence probe. Scale bars: 20 μm (a). Cellular ROS and mitochondrial ROS were stained with CellROX (green) and MitoSOX (red), respectively. Scale bars: 60 μm (b). Mn-SOD expression was detected by Western blot analysis (c). Protein lysates were incubated with DNPH to label carbonyl groups, and protein carbonylation was detected by Western blot analysis using anti-DNP antibodies (d). *P < 0.05, significantly different as indicated. ns Not significant. Biological replicates were performed for six times in the above studies