Anti pkcβ antibody

The pan anti-phospho-PKC activation loop antibody was described previously ^{107 (link)}. The anti-phospho-PKCα/βII (T638/641; 9375S) and pan anti-phospho-PKC (βII S660; 9371S) antibodies were purchased from Cell Signaling. Anti-PKCβ antibody was purchased from BD Transduction Laboratories. The anti-α-tubulin (T6074) antibody was from Sigma. Phorbol 12,13-dibutyrate (PDBu) and Uridine-5-triphosphate (UTP) were purchased from Sigma-Aldrich. 6His-N-terminally tagged human PLK4 (amino acids 1-269) was expressed and purified as described previously ^{108 (link)} and affinity purified using Ni-NTA agarose. Proteins were eluted from beads by incubation with 0.5 M imidazole. Described PLK4 mutations were introduced using standard PCR-based site-directed mutagenesis protocols and confirmed by sequencing the PLK4 1-269 coding region. pT170 PLK4 phosphospecific antibody was raised in rabbits against a phosphorylated PLK4 T-loop consensus peptide, and purified using standard affinity protocols prior to storage at -20°C ^{59 (link)}. The PLK4 inhibitors VX680 and staurosporine and the PLK1 inhibitor BI2536 were employed after dilution from 50 mM DMSO stocks stored at -80°C ^{79 (link)}.