Fitness and NPC clustering analysis of the Nup84 complex truncations. (A) Growth tests at different temperatures (24, 30, and 37°C) for the full-length (wild type [wt]) and truncated versions of the Nup84 complex nups. Serial 10-fold dilutions of cells were spotted on YEPD plates and grown at the indicated temperatures for 1–3 d. Parental strains DF5 and w303 as well as the full-length genomically tagged Nup84 complex nups (Nup84, Nup85, Nup133, Nup120, and Nup145c) were included as controls. Each growing phenotype was quantified by semiquantitative methods (see Materials and methods), and the obtained value (in arbitrary units [AU]) is shown on the right of each column. Plotted fitness value (mean ± SEM) for each measurement is shown on the right. (B) The left image of each column shows the localization of a Nup49p-CFP reporter in wild-type and truncation mutants as determined by fluorescence microscopy. A NUP133 gene deletion is also shown as a reference for NPC clustering (Belgareh and Doye, 1997). The right image of each column shows the differential interference contrast image of the same cells. The number shown on the bottom left corner of the fluorescence picture represents the measured degree of clustering for each strain (represented by their normalized CV, multiplied by 100 for representation purposes) in arbitrary units for n = 30 cells (see Materials and methods). Bars, 5 µm.