Figure 2. ima1Δ cells have NE and MAS defects.

A. Large deformations of the NE in the absence of Ima1. Fluorescent micrographs of one WT (top panels) and one ima1Δ cell (bottom panels) expressing Cut11-GFP and Sad1-DsRed (MKSP50 and MKSP56, respectively) taken in consecutive 60 second intervals are shown along with the merged images. Deformations led by Sad1-DsRed are labeled with arrows; deformations without visible Sad1-DsRed are labeled with asterisks. B. Large deformations of the NE in ima1Δ cells are driven by MTs. Fluorescent micrographs of one ima1Δ cell expressing GFP-tubulin and Heh1-mCherry (MKSP173) imaged every 30 seconds for one minute (time indicated on the left) and shown with the merged images. C. SPB oscillations are larger in ima1Δ cells. Composite images of time-lapse frames taken every 15 seconds for 5 minutes of one WT cell (MKSP81) and one ima1Δ cell (MKSP152) expressing GFPKms2 (upper panel). Plot of the maximum SPB oscillation amplitude for size-matched WT and ima1Δ cells (n=25, lower panel). The average value (ave) is given with its standard deviation. D. MT forces lead to defects in nuclear shape and cytoplasmic Cut11-GFP foci appear (arrows) in ima1Δ cells. Quantitation of the spherical nature of WT (MKSP10) versus ima1Δ (MKSP22) nuclei. As indicated, WT and ima1Δ cells were also treated for 15 minutes with carbendazim (MBC) prior to imaging. Measurements are presented from at least 100 non-mitotic cells. E. Percentage of WT (MKSP50) and ima1Δ (MKSP56) cells that display Cut11-GFP and Sad1-DsRed cytoplasmic foci. n = >100 interphase cells.