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. 2009 Jan 26;184(2):225–239. doi: 10.1083/jcb.200808049

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© 2009 Rogers et al.

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Figure 4. — Slimb regulates Plk4 levels on centrioles to control centriole number. (A) Asymmetrical Plk4 localization. Transient coexpression of Nlp-EGFP (Ito et al., 1996) as a cotransfection marker (green) labeling nuclei (arrow) and Plk4-EGFP (green). D-PLP (red) marks centrioles (arrowheads). Insets show centrioles at a higher magnification. (B–D) Mutating the Slimb-binding site stabilizes Plk4 on centrioles. (B) Cell cycle distributions after 24-h drug-induced S, G2, or mitotic arrest. Histograms are shown to scale and were assessed by HTM (5,000 cells/histogram). (C) Plk4-EGFP (green) only localizes to M-phase centrioles (arrowheads and insets) marked with mCherry–SAS-6 in live cells (red). Condensed DNA (blue) reveals mitotic cells. (D) Plk4-SBM–EGFP (green) localizes to centrioles during all cell cycle phases that were examined. Centrioles (arrowheads and insets) are marked with D-PLP in fixed cells (red). Nlp-EGFP (green nuclei, cytoplasmic during mitosis) is the cotransfection control. Bars: (A) 5 µm; (C and D) 2.5 µm.