Figure 2

KIF5B drives peripheral mitochondrial network formation. (A) CRISPR/Cas-mediated knockout of KIF5B in NRK cells stably expressing Mito-YFP. Expression levels of KIF5B were examined by western blotting. (B) Control and KIF5B-knockout cells were observed by confocal microscopy. Yellow dashed lines indicate the cell boundary; red dashed lines indicate the boundary of the mitochondrial network; and black dashed lines indicate the boundary of nucleus. Scale bar, 10 μm. (C) The cytoplasmic area occupied by mitochondria was quantified in n = 90 cells from three independent experiments. Error bars indicate SD. (D) KIF5B^−/− NRK cells that stably express Mito-YFP and TET-ON-KIF5B were treated with 0.5 μg/ml tetracycline. KIF5B induction was monitored by western blotting. (E) KIF5B^−/− NRK cells that stably express Mito-YFP and TET-ON-KIF5B T92N were treated with 0.5 μg/ml tetracycline. KIF5B T92N induction was monitored by western blotting. (F) Quantification of the area occupied by mitochondria in KIF5B^−/− NRK cells, TET-ON-KIF5B T92N cells, TET-ON-KIF5B cells, and TET-ON-KIF5B cells pretreated with 5 μg/ml nocodazole. n = 90 cells from three independent experiments. Error bars indicate SD.