Depletion of mDia2 produces a prolonged HJURP localization at centromeres. (A) Scheme for the siRNA transfection and the thymidine arrest to examine centromere localization of HJURP in early G1 cells. (B) Immunofluorescence images of GFP-HJURP. Centromeres were identified using ACA antibodies. The arrowheads point to centromeres shown in insets. Bars, 5 µm (insets are 2× magnified). (C) Whisker-Tukey boxplots show the relative intensity of GFP-HJURP foci at centromeres. The boxes span the 25th to 75th percentile of the samples, whereas the center bar denotes the median and the + marks the mean. Control: n = 174 centromeres from 26 cells, and mDia2 siRNA: n = 183 centromeres from 20 cells. The p-value was computed using a two-tailed t test. (D) Mean percentage of GFP-HJURP centromere-positive cells with error bars showing standard deviations from three experiments (control: n = 594 cells, and mDia2 siRNA: n = 277 cells). The p-value was computed using a two-tailed z-test. (E) Model of mDia2 regulating CENP-A loading. (left) Timeline of the epigenetic inheritance of CENP-A over cell cycle. (right) Schematic model showing mDia2 to be important for HJURP-mediated CENP-A chromatin assembly and timely HJURP turnover. Green, histone H3; red, old CENP-A; yellow, new CENP-A.