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. 2021 Jan 25;220(3):e202005153. doi: 10.1083/jcb.202005153

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© 2021 Ito et al.

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Figure 7. — Precocious centriole disengagement results in an increase of the number of centrioles. (A) Long-term Cep57/Cep57L1 codepletion increased the number of centrioles. HeLa cells were treated with siCep57/Cep57L1 for 96 h and immunostained with antibodies against centrin (green) and Cep192 (red). (B) Histograms represent the frequency of mitotic cells with the indicated number of centrioles observed in A. Values are mean percentages ± SD from three independent experiments (n = 30 for each experiment). Note that depletion of Cep57L1 for 96 h slightly reduced the number of centrioles. (C) HsSAS-6 was absent from disengaged centrioles and was reacquired. HeLa cells were treated with siControl or siCep57/Cep57L1 and immunostained with antibodies against HsSAS-6 (green) and CP110 (red). The insets in a–d are the magnified views of the corresponding regions in the low-magnification view. (D) Histograms represent the frequency of the number of HsSAS-6 foci on the two pairs of centrioles or four centrosomes in C. Values are mean percentages ± SD from three independent experiments (n = 30 for each experiment). (E) Precocious centriole disengagement and centriole reduplication in interphase and inevitable inheritance of amplified centrioles to daughter cells observed in Cep57/Cep57L1-depleted cells. HeLa-GFP-centrin1 (green) cells were treated with siCep57/Cep57L1. White arrowheads indicate centrosomes. (F) Histograms represent the frequency of mitotic cells with the indicated number of centrioles 48 h or 96 h after Cep57/Cep57L1 codepletion observed in A and in Fig. S5 B. Values are mean percentages ± SD from three independent experiments (n = 30 for each experiment). All scale bars, 5 µm in the low-magnification view, 1 µm in the inset. Dunnett’s multiple comparisons test was used in B to obtain the P value. ***, P < 0.001.