Figure 2. Ndel1 phosphorylation alters Lis1 and dynein interactions. (A) Schematic of five S/T-P sites in Ndel1 showing the residue recognized by the pS231Ab antibody. (B) Ndel1 was exposed to GST-cdk5/p25, GST-cdk1/cyclin B, or both. Top, Coomassie staining; middle, Ndel1 antibody; bottom, pS231Ab. (C, left) Immunoprecipitations of unphosphorylated or phosphorylated Ndel1 (Ndel1 and pNdel1) incubated with purified Lis1. (right) Quantification is representative of three experiments. (D) COS-7 cells were transfected with Ndel1, cdk5/p25, and cdk1/cyclin B or with Ndel15A, dncdk5, and dncdk1. (left) Immunoprecipitations for Ndel1 constructs. (right) Quantification is representative of three experiments. (E) Dynein co-IPs. (top) Probed for DIC, Lis1, and Ndel1. (bottom) Quantification is representative of three experiments. (F) Dynein co-IP with Ndel1 phosphorylated by individual kinases. LE, longer exposure.
Figure 2.

Ndel1 phosphorylation alters Lis1 and dynein interactions. (A) Schematic of five S/T-P sites in Ndel1 showing the residue recognized by the pS231Ab antibody. (B) Ndel1 was exposed to GST-cdk5/p25, GST-cdk1/cyclin B, or both. Top, Coomassie staining; middle, Ndel1 antibody; bottom, pS231Ab. (C, left) Immunoprecipitations of unphosphorylated or phosphorylated Ndel1 (Ndel1 and pNdel1) incubated with purified Lis1. (right) Quantification is representative of three experiments. (D) COS-7 cells were transfected with Ndel1, cdk5/p25, and cdk1/cyclin B or with Ndel15A, dncdk5, and dncdk1. (left) Immunoprecipitations for Ndel1 constructs. (right) Quantification is representative of three experiments. (E) Dynein co-IPs. (top) Probed for DIC, Lis1, and Ndel1. (bottom) Quantification is representative of three experiments. (F) Dynein co-IP with Ndel1 phosphorylated by individual kinases. LE, longer exposure.

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