Figure 1: EB2 is phosphorylated and not localized along MTs during mitosis.
From: Phosphorylation of EB2 by Aurora B and CDK1 ensures mitotic progression and genome stability
(a) Representative confocal images showing the localization of EB1 and EB2 throughout the cell cycle. Cells were co-stained with antibodies against EB1 (red), EB2 (green) and DAPI (blue). Scale bars, 5 μm. (b) HeLa cells were synchronized by a double-thymidine block and released into fresh medium. See Supplementary Fig. 9a for experimental scheme. Immunoblot analysis was carried out using antibodies against the indicated proteins. (c) In vitro CDK1 kinase assay. Left, active CDK1–Cyclin B1 was incubated with the indicated recombinant proteins in the presence of [γ-32P]-ATP. Right, the total amount of recombinant proteins from the same reaction were resolved by SDS–PAGE and Coomassie Brilliant Blue (CBB) staining. (d) HEK293T cells were co-transfected with EB2-HA in combination with GFP–Cyclin B1 and constitutively active FLAG-CDK1 (AF) or kinase-dead FLAG-CDK1 (DN), and incubated for 24 h. Immunoblot analysis was carried out using antibodies against the indicated tags.
