Fig. 6.

LATS and Aurora-B kinases phosphorylate Ser894 of INCENP in vivo. LATS2-knockout HeLa-S3 cells were transfected with siRNAs targeting LATS1 (siLATS1) and INCENP (siINCENP), and then treated with nocodazole (lanes 1–5 and 11–15) and taxol (lanes 6–10 and 16–20) in the presence or absence of Hesperadin (Hesp). siControl is a universal scrambled negative control siRNA duplex. The intensity of the pS894-specific band was reduced by knockout of LATS2 alone, and further reduced by knockdown of LATS1 or INCENP in LATS2-knockout cells (arrowhead in upper panels). The level of pS894 was normalized against the level of α-tubulin or INCENP. A double asterisk shows the degradation products of pS894-INCENP. These intensities were substantially reduced by additional treatment with Hesperadin (lanes 11–20). The slower-migrating bands of INCENP were converted to faster bands upon depletion of LATS1 and LATS2 (second panels from top). See Fig. S3 for uncropped western blot images in Fig. 6. Black boxes indicate the cropped regions. Molecular sizes (kDa) are based on prestained protein markers.