Figure S5.
The ciliary remnant in mitotic Nek2 KO cells can be rescued upon siRNA of Cep164 and expression of Nek2 in the Nek2 KO background.(A) Quantification of mitotic cells with Arl13B remnants at centrosomes in RPE1 Nek2 KO cells expressing DOX-inducible mNeonGreen-Nek2A. Cells were analyzed in the presence or absence of DOX. Bar graphs represent mean ± SD. n = 306 and 292 mitotic cells were analyzed without and with DOX treatment, respectively in three independent experiments. (B) Quantification of mitotic cells with Arl13B remnants at centrosomes in RPE1 WT and Nek2 KO cells treated with control (Ctrl) or Cep164 siRNA as indicated. The graph shows the average and SD of three independent experiments. The graph represents mean ± SD of three independent experiments. Numbers below the bars represent the total number of cells analyzed for each condition. (C) Boxplots represent the quantification of Cep164 appendage signal at the centrosome in RPE1 WT and Nek2 KO cells treated with control (Ctrl) or Cep164 siRNA as indicated. Data from three independent experiments are shown. Boxes show interquartile range, lines inside the box represent the median, and whiskers show minimum and maximum values excluding outliers. n  =  150 cells were analyzed for each condition. (D) Ciliation was examined in RPE1 WT and Nek2 KO cells after 48 h of serum starvation. Results show the mean ± SD of three independent experiments. Results show the average and SD of three independent experiments. Numbers below the bars represent the total number of cells analyzed for each condition. Representative images are shown on the right. Arl13B was used as a cilia and γ-tubublin (γ-tub) as a basal body marker. DNA was stained with DAPI. Scale bar, 10 µm. A.U., arbitrary units; n.s., not significant. Significance probability values are: n.s., P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.

The ciliary remnant in mitotic Nek2 KO cells can be rescued upon siRNA of Cep164 and expression of Nek2 in the Nek2 KO background. (A) Quantification of mitotic cells with Arl13B remnants at centrosomes in RPE1 Nek2 KO cells expressing DOX-inducible mNeonGreen-Nek2A. Cells were analyzed in the presence or absence of DOX. Bar graphs represent mean ± SD. n = 306 and 292 mitotic cells were analyzed without and with DOX treatment, respectively in three independent experiments. (B) Quantification of mitotic cells with Arl13B remnants at centrosomes in RPE1 WT and Nek2 KO cells treated with control (Ctrl) or Cep164 siRNA as indicated. The graph shows the average and SD of three independent experiments. The graph represents mean ± SD of three independent experiments. Numbers below the bars represent the total number of cells analyzed for each condition. (C) Boxplots represent the quantification of Cep164 appendage signal at the centrosome in RPE1 WT and Nek2 KO cells treated with control (Ctrl) or Cep164 siRNA as indicated. Data from three independent experiments are shown. Boxes show interquartile range, lines inside the box represent the median, and whiskers show minimum and maximum values excluding outliers. n  =  150 cells were analyzed for each condition. (D) Ciliation was examined in RPE1 WT and Nek2 KO cells after 48 h of serum starvation. Results show the mean ± SD of three independent experiments. Results show the average and SD of three independent experiments. Numbers below the bars represent the total number of cells analyzed for each condition. Representative images are shown on the right. Arl13B was used as a cilia and γ-tubublin (γ-tub) as a basal body marker. DNA was stained with DAPI. Scale bar, 10 µm. A.U., arbitrary units; n.s., not significant. Significance probability values are: n.s., P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.

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