Fig 2: Assembly of a pericentriolar kinase complex. a HEK293 cells were serum-deprived for 36 h and then left untreated (CNT) or stimulated for 3 h with isoproterenol (Iso) or forskolin (FSK) and doubly stained for acethylated tubulin and Draq5. Cumulative data from five independent experiments are shown. *p < 0.05 (Student's t-test). Where indicated, the cells were pretreated for 30 min with the PKA inhibitor (PKAi) H89 (5 µM) before stimulation. b RIIß was immunopurified from whole-cell extracts (WCE). IgG was used as control. The precipitates and lysates were immunoblotted with anti-PCM1 and anti-RIIß antibodies. c Lysates of NEK10-flag expressing cells were subjected to pull down assays with purified GST or GST–RIIß fusion. The precipitates and lysates were immunoblotted with anti-PCM1, anti-flag and anti-RIIß. d HEK293 cells were fixed and immunostained for endogenous PCM1, NEK10, and RIIß. A merge composite of the three signals and magnification of a selected area are shown (right panels)