Figure 1. Specificity of anti-(human INCENP) antibodies (α-INCENP) and depletion of INCENP or MKLP1 expression by corresponding siRNA in HeLa cells.

(A) Nocodazole-treated HeLa cells were lysed in lysis buffer, and cell lysates were immunoprecipitated with pre-immune antisera or anti-INCENP antibodies. Immunoprecipitates and one-tenth of whole-cell lysates (WCL) used in immunoprecipitations were subjected to SDS/PAGE (7.5% polyacrylamide), transferred on to a PVDF membrane and immunoblotted with anti-INCENP antibodies. M_r sizes are shown (×1000). (B) HeLa cells grown on six-well plates were transfected with control siRNA (120 nM) or INCENP siRNA (60–240 nM). At 3 days after transfection, cells were lysed in lysis buffer. Cell lysates were subjected to SDS/PAGE (7.5% polyacrylamide), transferred on to a PVDF membrane and then immunoblotted with anti-INCENP or anti-α-tubulin antibodies. (C) HeLa cells were transfected with 120 nM control siRNA, INCENP siRNA or MKLP1 siRNA. At 3 days after transfection, cells were lysed in lysis buffer. Cell lysates were subjected to SDS/PAGE (7.5% polyacrylamide), transferred on to a PVDF membrane and then immunoblotted with antibodies as indicated.