Fig. 9: Mitochondrial alterations caused by 181-Rik ablation or knockdown.

A BN-PAGE (blue native polyacrylamide gel electrophoresis) visualizes the integrity of the indicated respiratory complexes (CI-V) in mitochondria of 181-Rik WT and KO mouse telencephalons. B Transmission electron microscopy of microglial mitochondria from WT and KO retinas at 2 days post-IR injury. Red asterisks denote mitochondria. Scale bars: 1 μm (left panels) and 0.5 μm (right panels). C–F Quantification of the mitochondrial area (C), aspect ratio (D), circularity (E), and crista area proportion (F). Data were presented as mean ± SD [n = 91 and 97 (C), 89 and 97 (D), 90 and 97 (E), and 57 and 88 (F) for WT and KO retinas, respectively]. G–I qRT-PCR assay of mitochondrial fusion-related genes Opa1 and Mfn1 and fission-related gene Mff (G), immunostaining for Mfn1 and Mff (H), and quantification of the corresponding immunofluorescence intensity (I) in microglia infected with 181-Rik shRNA or scramble lentiviruses. Data were presented as mean ± SD [n = 5 or 6 (G) and 6–8 (I)]. AU arbitrary unit. Scale bar: 25 μm. J–L DHE (dihydroethidium) labeling (J), quantification of the corresponding immunofluorescence (representing ROS) intensity (K), and MMP (mitochondrial membrane potential) measurements (L) in the four groups of microglia (scramble, shRNA, scramble+OGD/R, and shRNA+OGD/R). Data were presented as mean ± SD [n = 8–10 (K) and 4 (L)] in arbitrary unit (AU). Scale bar: 100 μm.