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. Author manuscript; available in PMC: 2015 Aug 21.
Published in final edited form as: Cell Rep. 2014 Aug 14;8(4):957–965. doi: 10.1016/j.celrep.2014.07.022

Figure 4. Centriole-to-centrosome conversion stabilizes centrioles devoid of the cartwheel.

Figure 4

(A) Mother centrioles and their engaged daughter centrioles in cells treated with control (left) or CEP295 siRNA (right) for 72 hours were examined at early mitosis with antibodies against centrin, γ-tubulin (gTub), and acetylated α-tubulin (AcTu).

(B) Cells treated with control or CEP295 siRNA for 72 hours were labeled with BrdU before fixation. G1 cells, BrdU negative (not shown) and inheriting a pair of disengaged centrioles, were examined with indicated antibodies. Quantifications are shown (right) (n>100, N=3). Error bars represent standard deviation. Note that in CEP295 siRNA-treated cells, majority of daughter centrioles failed to convert to centrosomes (group II & III; gTub-), some of which also lost the AcTu staining (group III) (gTub- & AcTu-).

(C) Cells were treated as described in (B). BrdU-positive cells inheriting active centrosomes (gTub foci) were examined with indicated antibodies. Cells inheriting no centrosomes were excluded. Quantifications are shown (right) (n>100, N=3). Note that in the majority of CEP295 RNAi cells, the inherited DCP/daughter centrioles either became non-detectable (group IV), or had lost the AcTu staining (group III) (gTub-; AcTu-).

(D) DCP centrioles in S-phase cells were examined for the localization of CPAP with indicated antibodies.

See also Figure S3.