Figure 3.
Overexpression of WDR62-GFP wild type and c.1313G>A ( p.Arg438His) mutant constructs in HeLa cells. The first 4.5 panel of images (on the left) shows the results for the wild-type WDR62-GFP construct, whereas the second 4 . 5 panel of images (on the right) shows the comparable results for the c.1313G>A (p.Arg438His) missense mutation in the WDR62-GFP construct. The wild-type WDR62-GFP protein localized to the spindle pole during mitosis (in 50 of 50 cells analyzed), paralleling the localization found for the endogenous wild type protein. Conversely, in cells expressing the mutant construct, there was no spindle pole accumulation of GFP during mitosis (in 50 of 50 cells analyzed). Cells were stained with γ-tubulin (red) as a centrosome marker and α-tubulin (white) identifying microtubules and the mitotic spindle. Both WDR62-GFP (green) proteins were directly visualized with DNA (blue) stained with DAPI. (See Supplementary Fig. 4 for the results for the c.4241dupT mutation). Scale bar, 5 μm.