Figure S3.
LIC1-CTD phosphorylation is required for prometaphase kinetochore dynein recruitment in HeLa cells. (A, C, E, and G) Representative confocal immunofluorescence micrographs of prometaphase HeLa cells (1 µM nocodazole treated for 4 h), immunostained to visualize the chromosomes (DAPI, blue), kinetochores (CREST, red), and the indicated kinetochore proteins (green) as follows: IC (A), spindly (C), Zw10 (E), and p150Glued (G). All cells were treated with anti-hLIC1 siRNA and rescued by the transient expression of the indicated rLIC1-CTD constructs. (B, D, F, and H) Scatterplots depicting the quantification of the immunofluorescence signals from A, C, E, and G, respectively, normalized to the respective kinetochore (CREST) intensities, upon rescue of hLIC1 depletion by the indicated rLIC1 constructs (x axis). A minimum of 80 prometaphase cells per condition, over 3 independent experiments, were quantified. Scale bar = 10 µm. Error bars = mean ± SD. ***, P < 0.001 (two-tailed Student’s t test).