Ndel1 negatively controls ciliary length in Swiss 3T3 cells. (A) Swiss 3T3 cells were treated with each Ndel1-specific siRNA (target sequence 3 or m1) or control siRNA for 48 or 72 h. Cell lysates were immunoblotting with anti-Ndel1 and anti-actin. (B) Tet-On Swiss 3T3 cell line expressing inducible GFP or Ndel1-Myc was incubated with (+) or without (−) 30 ng/ml Dox in the growing medium for 24 h and then in the medium containing 0.1% serum for 24 h. Cell lysates were immunoblotting with anti-Myc, anti-GFP, and anti-actin. The data are shown as mean ± SEM from three independent experiments (n > 100 each; A, B, and C–E) Swiss 3T3 cells were treated with the indicated siRNA. 24 h after transfection, the medium was changed to new medium containing 0.1% serum. Then, the cells were incubated for additional 48 h. Averaged (C) and distribution (D) of ciliary length are shown in the graphs. Ciliary length from 40 cells was measured for each experiment (means ± SEM; n = 3 independent experiments). The treated cells were stained with anti–acetylated tubulin (ac-tub.; green), anti-Ndel1 (red), and DAPI (blue; E). *, P < 0.05; ***, P < 0.001; n.s., not significant; two-tailed unpaired t test. Bars: (E, bottom) 5 µm; (E, top) 10 µm.