Supplemental Materials

This article contains the following supporting material:

• Figure S1 - Mapping of the pericentrin-binding domain on CHD4 using the yeast two-hybrid system
  A series of GAL4-TAD CHD4 deletion constructs including the pericentrin-interacting fragment identified in the yeast two-hybrid screen and a GAL4-DBD pericentrin expression construct were used to transform the yeast strain AH109. A diagram outlining the N-terminal (clear boxes) and C-terminal (black boxes) deletion mutants used is presented in the center of the figure. Images of yeast growing on selective media are shown with the left quadrant of the plate being a DBD x TAD-CHD4 control and the right quadrant a DBD-pericentrin x TAD-CHD4 experimental. C-terminal deletions are on the left and N-terminal deletions are on the right. Two regions between residues 1577-1733 and 1783-1912 of CHD4 were found to be able to interact with pericentrin. Sequence comparison shows the C-terminus of CHD4, residues 1577-1912, is 59% identical and 72% similar to the C-terminus of CHD3.
• Figure S2 - Disrupted spindles in CHD3 depleted cells
  Forty-eight hours after CHD3 depletion, HeLa cells were stained with anti-α-tubulin (microtubules, red) and 5051 antibodies (centrosomes, green). Insets show enlargements of centrosome staining. All spindles show reduced microtubule numbers. Scale bars 10 μm.
• Figure S3 - Delay in microtubule regrowth in CHD3 siRNA-treated cells after nocodazole washout
  Additional images of regrowing microtubules in CHD3 siRNA-treated cells fixed 5 minutes after nocodazole washout. The CHD3 depleted cell in the upper left panel has fewer microtubules associated with centrosomes (in boxes), many non-centrosome-associated microtubules in the periphery, a large microtubule free region surrounding asters and a nucleus displaced from the cell center. Scale bars μm.
• Movie S1
• Movie S2