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. 2014 May 22;111(23):8541–8546. doi: 10.1073/pnas.1323161111

Fig. 2.

Fig. 2.

ICK kinase activity levels determine the length of primary cilia. (A and B) FLAG-tagged wild-type and kinase-inactive forms of ICK (K33R and TDY mut) were transfected into cultured cells. (A) Cilia and transfected ICK were visualized by immunofluorescent staining with acetylated tubulin (acTub, green) and FLAG (red) antibodies, respectively. Overexpression of wild-type ICK either suppressed cilia formation or shortened ciliary length. By contrast, overexpression of inactive ICK increased ciliary length. (B) Quantification of ciliary length in CTRL (n = 56), K33R (n = 56), or TDY mut (n = 58) ICK overexpressing cells. (C) Western blot analysis showed that ICK protein levels were significantly decreased by Ick siRNA 2 compared with control siRNA or siRNA 4. (D and E) Primary cilia were stained by Arl13b antibody (green) to analyze cilia length (n = 14) (D) and morphology (E). Knockdown of ICK by siRNA 2 resulted in longer primary cilia than those observed after control siRNA or siRNA 4 treatment. Error bars reflect standard SEM and asterisk denotes statistical significance according to Student’s t tests in B and D (*P < 0.01). (Scale bars in A and E, 5 µm.)