GST-Ana2-S38A can bind Sas6 after Plk4 phosphorylation from Two-step phosphorylation of Ana2 by Plk4 is required for the sequential loading of Ana2 and Sas6 to initiate procentriole formation

Supplementary Figure S3: GST-Ana2-S38A can bind Sas6 after Plk4 phosphorylation. Sas6 specifically interacts with both Ana2-WT and Ana2-S38A when the proteins are pre-phosphorylated by Plk4. GST, GST-tagged wild-type (GST-Ana2-WT) or the S38A substitution mutant (GST-Ana2-S38A) were treated with either MBP-Plk4 or MBP-Plk4KD and incubated in vitro with 35S-Met-labeled Sas6 produced in a coupled in vitro transcription/translation reaction. The resulting complex was analyzed by SDS-PAGE and autoradiography.