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. 2019 Aug 15;2(4):e201900308. doi: 10.26508/lsa.201900308

Figure S1. Generation of brain-specific MITOL KO mice.

Figure S1.

(A) Schematic representation of the strategy for gene targeting of march5/mitol. The genomic structure of the WT march5/mitol allele (top) and the march5/mitol allele (bottom) are shown. Targeting strategy for march5/mitol gene: homologous recombination of the targeting vector and generation of the march5/mitol KO by activation of the Cre-loxP system. Triangles, loxP sites; Neo, neomycin resistance gene. (B) Genome PCR confirmed march5/mitol genome deletion. (C) Immunoblot analysis indicates the loss of MITOL expressions in cerebral cortex but not cerebellum of eKO. (D, E) No obvious differences in both body size and brain size between WT and eKO. (F) A coronal section of cerebrum obtained from 3-mo-old WT. Samples including corpus callosa (CC) and hippocampi (*) were collected from the area marked with a box. (G) A 1 μm-thick section of a metal-stained and resin-embedded tissue sample. Serial electron microscopic images were obtained form the border of pyramidal cell layer (py) and stratum oriens (or) in hippocampal CA1 for neuronal cell bodies (b, black box). Scale bar represents 100 μm. (H) All mitochondria showed in scatter plot of mitochondrial volume versus sphericity. (n = 225 for mitochondria in cell body). (I) Comparison of activated form of Drp1. Cerebral cortex and hippocampus (CH) or ceberellum (Cb) of WT or eKO were analyzed by IB with the indicated antibodies. Co, Cortex; DG, Dentate gyrus; ra, Stratum radiatum.