Figure 9.
Effect of Kif24 upon appendages.(A) RPE1 mNeonGreen-Nek2A (Nek2 WT) cells were treated with control (siCtrl) or Kif24 siRNA before addition of solvent (− DOX) or doxycycline (+ DOX). Box/dot plots show quantification of the indicated appendage intensities at centrosomes. Boxplots show fluorescence intensity (int.) measurements for Cep164 in interphase cells. Boxes show interquartile range, lines inside the box represent the median, and whiskers show minimum and maximum values excluding outliers. Data of three independent experiments are shown. n = 175 cells per condition. (B) qPCR was used to quantify Kif24 mRNA levels in RPE1 cells treated with control or Kif24 siRNAs. Results show the RQ value for the respective treatment, and error bars show the respective maximum and minimum RQ value for each sample. One representative experiment of two is shown. (C) Representative images show Cep164 staining in interphase and mitotic RPE1 cells treated with control or Kif24 siRNA. The insets show magnification of the centrosome area. Scale bars, 10 µm (panel) and 2 µm (inset). γ-tub, γ-tubulin. (D) Western blot shows AcGFP1-Kif24 levels using anti-GFP antibodies. Cofilin was used as a loading control. (E) Analysis of Cep164 upon AcGFP1-Kif24 expression under the strong cytomegalovirus promoter. Fluorescence intensity (int.) measurement of Cep164 in control (Ctrl) and AcGFP1-Kif24–transfected RPE1 cells. Boxes show interquartile range, lines inside the box represent the median, and whiskers show minimum and maximum values excluding outliers. Data from three independent experiments are shown. n = 150 per condition. (F) Representative images show AcGFP1-Kif24 and Cep164 centrosomal staining in control (Ctrl) and AcGFP1-Kif24–transfected cells. The insets show enlargements of the centrosomal area. Scale bar, 10 µm (panel) and 2 µm (inset). (G) Ciliation was quantified in control (Ctrl) and AcGFP1-Kif24–transfected RPE1 cells. Cells were transfected with AcGFP1-Kif24 for 48 h and serum starved for 24 h and stained with anti-GFP and anti-Cep164 antibodies. n = 62 (Ctrl) and n = 29 (AcGFP1-Kif24). n.s., not significant. A.U., arbitrary units. Significance probability values are: n.s., P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.

Effect of Kif24 upon appendages. (A) RPE1 mNeonGreen-Nek2A (Nek2 WT) cells were treated with control (siCtrl) or Kif24 siRNA before addition of solvent (− DOX) or doxycycline (+ DOX). Box/dot plots show quantification of the indicated appendage intensities at centrosomes. Boxplots show fluorescence intensity (int.) measurements for Cep164 in interphase cells. Boxes show interquartile range, lines inside the box represent the median, and whiskers show minimum and maximum values excluding outliers. Data of three independent experiments are shown. n = 175 cells per condition. (B) qPCR was used to quantify Kif24 mRNA levels in RPE1 cells treated with control or Kif24 siRNAs. Results show the RQ value for the respective treatment, and error bars show the respective maximum and minimum RQ value for each sample. One representative experiment of two is shown. (C) Representative images show Cep164 staining in interphase and mitotic RPE1 cells treated with control or Kif24 siRNA. The insets show magnification of the centrosome area. Scale bars, 10 µm (panel) and 2 µm (inset). γ-tub, γ-tubulin. (D) Western blot shows AcGFP1-Kif24 levels using anti-GFP antibodies. Cofilin was used as a loading control. (E) Analysis of Cep164 upon AcGFP1-Kif24 expression under the strong cytomegalovirus promoter. Fluorescence intensity (int.) measurement of Cep164 in control (Ctrl) and AcGFP1-Kif24–transfected RPE1 cells. Boxes show interquartile range, lines inside the box represent the median, and whiskers show minimum and maximum values excluding outliers. Data from three independent experiments are shown. n = 150 per condition. (F) Representative images show AcGFP1-Kif24 and Cep164 centrosomal staining in control (Ctrl) and AcGFP1-Kif24–transfected cells. The insets show enlargements of the centrosomal area. Scale bar, 10 µm (panel) and 2 µm (inset). (G) Ciliation was quantified in control (Ctrl) and AcGFP1-Kif24–transfected RPE1 cells. Cells were transfected with AcGFP1-Kif24 for 48 h and serum starved for 24 h and stained with anti-GFP and anti-Cep164 antibodies. n = 62 (Ctrl) and n = 29 (AcGFP1-Kif24). n.s., not significant. A.U., arbitrary units. Significance probability values are: n.s., P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal