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. 2004 Dec;24(24):10621–10635. doi: 10.1128/MCB.24.24.10621-10635.2004

FIG. 4.

FIG. 4.

Helix mutations perturb tight ring formation in early mitosis. (A) Localization of mid1-GFP (lane a, AP528), NLS* mid1-GFP (lane b, AP734), Helix* mid1-GFP (lane c, AP583), and Helix* NLS* mid1-GFP (lane d, AP630) in cells grown at 20°C. mid1-GFP and NLS*mid1-GFP form a broad band at the medial cortex during interphase and assemble a medial tight ring during mitosis (arrowheads). Helix* mid1-GFP is nuclear in interphase, whereas Helix* NLS* mid1-GFP is mostly cytoplasmic but is also present in the nucleus and forms faint patches at the cell cortex (long arrows). Both form disorganized filaments in the medial region in early mitosis (short arrows). Bar, 5 μm. (B) Expression levels of mid1-GFP (a), NLS* mid1-GFP (b), Helix* mid1-GFP (c), and Helix* NLS* mid1-GFP (d) at 30°C and 36°C. Western blot with anti-mid1p antibody and MAb TAT1 for tubulin (loading control). WT, wild-type cells.