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. 2019 Jul 29;10(8):570. doi: 10.1038/s41419-019-1778-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Alignment of amino acids in human Cep131 and several other species. The phosphorylation sites at residues S21 by Plk4 are highlighted in red. Asterisk, fully conserved residue; colon, conserved residue; period, semi-conserved residue. b Peptide competition assay to verify the specificity of anti-Cep131-pS21 antibody. Lysates from HEK293T cells expressing Myc-Plk4 with HA-Cep131-WT or S21A were incubated with non- or phosphopeptides targeting Cep131-S21. After incubation, phosphorylation status was analyzed by immunoblotting using Cep131-pS21 antibody. c HEK293T cells co-expressing HA-Cep131-WT with Myc-Plk4-WT or KD were immunoprecipitated using HA-magnetic beads, followed by incubation with λ-phosphatase at 30 °C for 30 min. Phosphorylation status was then observed by immunoblotting using anti-Cep131-pS21 antibody. d Immunofluorescence analysis of co-localization of phosphorylated Cep131 at the centriole. U2OS cells were treated with shGL2 (control) or shCep131 and were co-stained for Cep131-pS21 (red) and Cent (green). e Quantification of Cep131-pS21-positive cells. Error bars represent means ± SEM from three independent experiments (N > 300 for each experiment). **P < 0.01, unpaired Student’s t test. CBB staining use as a loading control in b, c