Fig. 8
Mis18V211D is a separation-of-function mutant with impaired HJURP binding but not M18BP1 binding. a Mutated residues of Mis18α and Mis18β are indicated on the 3D model. b, c SDS-PAGE results of the amylose-resin pull-down assays for testing the interaction of the mutants of Mis18α and Mis18β with HJURP394–C, M18BP11–60, and M18BP161–140. d Western blotting analysis confirming transgene expression. Endo. endogenous. e Representative images showing the fluorescence of mCherry-HJURP and EGFP-Mis18α variants in fixed HeLa cells treated as described in Fig. 2a but without the blocking and labeling steps of SNAP-CENP-A. Mis18α siRNA was used instead of HJURP siRNA. White scale bars indicate 10 µm. f, g Quantification of the centromere fluorescence intensity of mCherry-HJURP and EGFP-Mis18α variants. The bar graphs represent mean values from three replicate experiments (blue dots indicate the mean values from each experiment). Error bars indicate standard deviations. h Two models of HJURP recruitment to the Mis18 receptor and the deposition of CENP-A:H4 dimers. Source data are provided as a Source Data file
