FG-nucleoporins Nup153, Nup214, and Nup358 serve different functions for the transport of MAD into the nucleus. (A) GFP-MAD+R cells were treated with the indicated RNAi and were induced to express GFP-MAD and Punt/Tkv. The distribution patterns of GFP-MAD (green) and DAPI (blue) are shown. Bar, 10 μM. (B) Quantification of the nuclear/cytoplasmic ratio of GFP-MAD signals in panel A. Data are means ± SEs for ≥3 fields. (C) GFP-MAD+R cells were treated with the indicated RNAi, and the cell extracts were analyzed by immunoblotting with MAb414 and antitubulin. (D) Quantification of the nuclear/cytoplasmic ratio of cNLS-GFP-MAD in cells treated with the indicated RNAi. Data are means ± SEs for ≥3 fields. (E) S2R+ cells were treated with the indicated RNAi, and the cells were double stained with MAb414 (green) and anti-Msk (red). Bar, 5 μM.