Stabilization of INCENP by Aurora-C phosphorylation.
(A) Flag-Aurora-C WT, KD, T198A, or T202A mutants were co-transfected with HA-INCENP WT or 3A mutant and GFP empty vector in 293T cells. After 24 h transfection, cells were analyzed. GFP was used as internal control for transfection. Quantification of HA-INCENP protein levels were determined by using the ImageJ software. Relative fold change with normalization by GFP expression is shown. (B) 24 h after transfection, 293T cells were split and grown in culture for additional 24 h. Then cells were treated with 50 μM LLnL for 6 h and analyzed as in A. Experiments (A) and (B) were repeated three times with two biological replicates and figures show the representative results.