JCS048223 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Interactions among Plk1, FAM29A and NEDD1. (A) HeLa S3 cells stably expressing GFP-S-Plk1 were synchronized by a thymidine-nocodazole arrest to enrich prometaphase cells. The Plk1 complexes were tandem-affinity-purified and the associated proteins were identified by mass spectrometry (Seki et al., 2008a; Seki et al., 2008b; Zhu et al., 2008). Peptides of FAM29A identified in the Plk1 complex are listed with XCorr and DeltaCN scores indicated. (B) Maximum projections from deconvolved z-stacks of representative HeLa cells stained for NEDD1 (green), Plk1 (red) and DNA (blue). Scale bars: 5 µm.

• Supplemental Figure S2 -

  Fig. S2. Treatment with BI 2536 or knockdown of Plk1 does not alter the expression levels of FAM29A and NEDD1. (A) HeLa cells were treated with DMSO and BI 2536 (300 nM) for 2 hours and protein levels were analyzed by western blotting. (B) HeLa cells were transfected with siRNAs and protein levels were analyzed by western blotting at 48 hours post-transfection. Knockdown of Plk1 arrests cells in mitosis. Thus, there is an increase in the hyperphosphorylated FAM29A and NEDD1 in the siPlk1 sample.