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. Author manuscript; available in PMC: 2020 Mar 31.
Published in final edited form as: Curr Biol. 2018 May 3;28(10):1643–1650.e3. doi: 10.1016/j.cub.2018.04.014

Figure 1: Nup98 is required for cilia length regulation.

Figure 1:

See also Figure S1. (A) siRNA-mediated knockdown of Nup98 in hTERT-RPE (RPE) cells results in a loss in the fluorescence intensity of Nup98 staining at the ciliary base, confirming antibody specificity. (B) siRNA-mediated knockdown results in a 75–80% reduction of Nup98 protein levels. The Western blot was quantified in ImageJ and repeated 2 more times with similar results (quantitation shown at upper edge of Western blot). (C) Fluorescence intensity of Nup98 was measured at the ciliary base using ImageJ, and normalized to the fluorescence intensity of the mother centriole marker, Ninein. There was a specific reduction in Nup98 signal in cells treated with a Nup98 siRNA, when compared to controls. (D) Cilia length was decreased in serum-starved RPE cells treated with a Nup98 siRNA. (E) After knockdown of Nup98, RPE cells no longer show cilium elongation in response to serum-starvation.

p values in (C, D) were calculated by unpaired t test. For (E) a one-way ANOVA was performed comparing cilia length before starvation to the 4h, 8h, and 24h time points during starvation. For control siRNA, p < 0.0001. For Nup98 siRNA, p = not significant. Cilia length was measured using ImageJ. In (C), error bars are S.D.; all other error bars are S.E.M.