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. 2018 Apr 2;217(4):1217–1231. doi: 10.1083/jcb.201605106

Figure 4.

Figure 4.

Centrioles containing Ana2-7A display an abnormal architecture but this is not caused by a failure to disengage. (A and B) The morphology of the centriole outer surface protein, PLP, is abnormal in Ana2-7A–expressing cells. Interphase centrioles in inducible GFP-Ana2-WT (A) or GFP-Ana2-7A (B) stable cell lines were imaged using SR-SIM. Expression of GFP-Ana2 (green) was induced for 5 d. Cells were immunostained for PLP (red) and Hoechst-stained for DNA (not depicted). The number of centrioles with normal (A and B, row 1) or abnormal (B, rows 2–4) morphology is indicated. Bars, 200 nm. (C) Graph shows mean lengths of the major axis of centrioles in interphase cells. Measurements are shown for disengaged single centrioles in DMSO-treated cells (n = 17), engaged centriole pairs in BI-2536–treated cells (n = 9), and centrioles in stable GFP-Ana2–expressing cells (WT, n = 19; 7A, n = 13; 7PM, n = 34). Asterisks mark significant differences between treatments: **, P < 0.01; ****, P < 0.0001. Error bars, SEM. (D) Chemical inhibition of Polo blocks centriole disengagement in S2 cells. Cells were treated for 48 h with either DMSO or the BI-2536 Polo inhibitor, and interphase centrioles were imaged using SR-SIM. Cells were immunostained for PLP (red) and Asl NT (blue). Bars, 200 nm. (E) Examples of centriole measurements in control and Polo-inhibited interphase S2 cells. Cells were treated and prepared for SR-SIM as in D. The major axis lengths of single centrioles and engaged pairs was measured. The numbers of measured single centrioles (top) and engaged pairs (bottom) are indicated. Bars, 200 nm.