Figure 3.
Depletion of CEP55 increases the percentage and length of primary cilia in cultured cells. (A) RPE-1 cells transfected with control or CEP55 siRNA in the presence of serum (+ Serum) were stained with Ac-tubulin (green), γ-tubulin (red), and DNA (blue). Insets show zoomed-in views of the boxed regions. Scale bars, 10 µm (main image) and 1 µm (magnified region). (B) Effects of CEP55 depletion on cilia formation in the presence of serum (+ Serum) in RPE-1 cells. Quantification of the ciliated cells in A with three individual siRNAs against CEP55. Data are means ± SD of three independent experiments. One-way ANOVA test was performed followed by Dunnett's multiple comparisons. ***, P < 0.001. n, number of cells. (C) Immunoblot of RPE-1 cells lysates in A and B with the indicated antibodies. (D) RPE-1 cells transfected with control or CEP55 siRNA were stained with Ac-tubulin (green), γ-tubulin (red), and DNA (blue) under serum starvation (− Serum). Insets show zoomed-in views of the boxed regions. Scale bars, 10 µm (main image) and 1 µm (magnified region). (E) Quantitative analysis of the cilium length in D with three individual siRNAs against CEP55. Each dot represents one cell. Data are means ± SD. A one-way ANOVA test was performed followed by Dunnett’s multiple comparisons. ***, P < 0.001. (F) RPE-1 cells were transfected with mCherry-vector (−), mCherry-CEP55-WT, or mCherry-CEP55 3SA and then starved for 48 h. Cells were stained with Ac-tubulin (green) and DNA (blue). Insets show zoomed-in views of the boxed regions. Scale bars, 5 µm (main image) and 1 µm (magnified region). (G) Quantification of the ciliation in mCherry-positive cells in F. Data are means ± SD of three independent experiments. A one-way ANOVA test was performed. ***, P < 0.001. n, number of cells. (H) Immunoblot of RPE-1 cells lysates in F and G with the indicated antibodies.

Depletion of CEP55 increases the percentage and length of primary cilia in cultured cells. (A) RPE-1 cells transfected with control or CEP55 siRNA in the presence of serum (+ Serum) were stained with Ac-tubulin (green), γ-tubulin (red), and DNA (blue). Insets show zoomed-in views of the boxed regions. Scale bars, 10 µm (main image) and 1 µm (magnified region). (B) Effects of CEP55 depletion on cilia formation in the presence of serum (+ Serum) in RPE-1 cells. Quantification of the ciliated cells in A with three individual siRNAs against CEP55. Data are means ± SD of three independent experiments. One-way ANOVA test was performed followed by Dunnett's multiple comparisons. ***, P < 0.001. n, number of cells. (C) Immunoblot of RPE-1 cells lysates in A and B with the indicated antibodies. (D) RPE-1 cells transfected with control or CEP55 siRNA were stained with Ac-tubulin (green), γ-tubulin (red), and DNA (blue) under serum starvation (− Serum). Insets show zoomed-in views of the boxed regions. Scale bars, 10 µm (main image) and 1 µm (magnified region). (E) Quantitative analysis of the cilium length in D with three individual siRNAs against CEP55. Each dot represents one cell. Data are means ± SD. A one-way ANOVA test was performed followed by Dunnett’s multiple comparisons. ***, P < 0.001. (F) RPE-1 cells were transfected with mCherry-vector (−), mCherry-CEP55-WT, or mCherry-CEP55 3SA and then starved for 48 h. Cells were stained with Ac-tubulin (green) and DNA (blue). Insets show zoomed-in views of the boxed regions. Scale bars, 5 µm (main image) and 1 µm (magnified region). (G) Quantification of the ciliation in mCherry-positive cells in F. Data are means ± SD of three independent experiments. A one-way ANOVA test was performed. ***, P < 0.001. n, number of cells. (H) Immunoblot of RPE-1 cells lysates in F and G with the indicated antibodies.

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