Fig. 2

NEK10 interacts with GLUD1. a Proximity ligation assay of NEK10 vs. GLUD1/CS. HeLa pLKO cells were incubated with NEK10 (mouse or goat), GLUD1 (goat) and CS (mouse), and incubated with anti-mouse plus and anti-goat minus PLA reagents, followed by ligation, amplification and far-red staining reagents; nuclei were stained with Hoechst 33342 reagent. b Quantitative analysis of the average number of NEK10 and GLUD1 (or CS) interaction, indicated as PLA spots per cell, is represented in the bar graph. The images were collected with the ImageXpress, Micro Confocal High Content Image System (Molecular Devices) and the data were obtained with the MetaXpress software by using the transfluor module. One way ANOVA was used followed by the Bonferroni post hoc test. *** = P < 0.001. c Colocalization of NEK10 and Glutamate Dehydrogenase- 1 (GLUD1). Confocal images of HeLa cells stained with ant-NEK10 (in green) and anti-GLUD1 (in red) showing colocalization (merge, yellow) in the perinuclear region (arrows). Cells were visualized using a confocal Zeiss LSM 510, with a 63x oil objective, 1 air unit pinhole, 1024 × 1024 pixels. Scale bars: 10 μm. Images were analyzed using FIJI software [26]