Fig. 3

NEK10 is localized in the mitochondria. a Mitochondrial fractionation assay. Mitochondria from HeLa pLKO, HeLa pLKO-sh89 and HeLa pLKO-sh90 cells were isolated and NEK10 was analyzed by Western blot using anti-NEK10 antibody. Postnuclear supernatant (PNS), cytosol (CYT) and mitochondrial (MITO) fractions were analyzed with anti-Tubulin B, anti-OXPHOS and anti-VDAC antibodies, for fraction purity. The letter A and the red arrows indicate the 133 kDa isoform, present in PNS and CYTO. The letter B and the blue arrows indicate the 80 kDa isoform, present in the mitochondrial fraction (MITO). b Confocal images of NEK10 in mitochondria. HeLa pLKO-empty cells were stained using anti-NEK10 antibody (green) and the mitochondria were visualized with MitoTracker DeepRed for 20 min (red). Cells were analyzed in a confocal Zeiss LMS 510 microscope, using 63x oil objective, 1 air unit pinhole, 2048 × 2048 pixels. Scale bars: 10 μm. Images were analyzed using FIJI software [26]. Insets are enlargements of the outlined areas