MCPH-associated mutations within WD40 domain of WDR62 disrupt its interactions with both MTs and Aurora A. (A) Immunofluorescence staining of α-tubulin in MRC5 cells transiently expressing GFP-tagged Aurora A FL, N-terminal part (NT), or C-terminal kinase domain (CT). All these Aurora A constructs by themselves failed to localize to MTs. (B) Images of fixed MRC5 cells transiently expressing WDR62 WD-mCherry together with GFP-tagged Aurora A FL, NT, or CT. WDR62 WD-mCherry can recruit GFP-tagged Aurora A FL and CT, but not NT, to MTs. (C) StrepTactin pull-down assays with extracts of HEK293T cells expressing Strep-GFP-Aurora A (bait) together with GFP-tagged WT WDR62 or its indicated MCPH-associated mutants (prey). (D and E) Immunofluorescence staining and quantification of katanin p80 intensities at spindle poles in WDR62 knockout (KO) HeLa cells transiently transfected with GFP-tagged WT WDR62 or its indicated MCPH-associated mutants. For all conditions, n = 42 spindle poles. (F) Immunofluorescence staining of α-tubulins in MRC5 cells transiently transfected with GFP-tagged WT WDR62 or its indicated MCPH-associated mutants. (G) Purified proteins used for in vitro assays. The bands corresponding to indicated proteins are marked with red asterisks on Coomassie blue–stained gels. Note that molecular chaperone HSP70 (∼70 KD) was copurified with all indicated proteins. Scale bars, 2 µm. Data represent mean ± SD.