NuSAP had no effect on the localization of KifC1 and MCAK. (A) NuSAP interacted with Kif2A, Eg5, KifC1, and MCAK. Mitotic HEK293T cells were transfected with GFP or GFP-NuSAP and then immunoprecipitated with GFP-Trap beads. (B and D) Sample images (top) and quantification (bottom) of relative KifC1 (B) and MCAK (D) fluorescence intensity in the control and NuSAP-knockout cells, respectively. Error bars indicate SD. Three independent replicates of 30 cells per replicate were quantified (n = 3). Unpaired two-tailed t test: P = 0.7074 in B; P = 0.7179 in D. The white dotted lines show the areas of measurement. (C and E) Sample images and quantification of relative KifC1 (C) and MCAK (E) fluorescence intensity in HeLa cells transfected with GFP or GFP-NuSAP, respectively. Error bars indicate SD. Three independent replicates of 30 cells per replicate were quantified (n = 3). Unpaired two-tailed t test: P = 0.6413 in C; P = 0.0880 in E. The white dotted lines show the areas of measurement. ns, not significant. n = the number of independent experiments presented. Scale bars, 10 μm. Source data are available for this figure: SourceData FS3.