Figure 3.
Effect of actin depolymerization by LatB on Kar9p-GFP3 localization. (A) After arrest in G1 by α factor, wild-type cells were released and allowed to proceed past bud emergence. At the point indicated by the arrow, the culture was divided for treatment with either 100 μM LatB or dimethyl sulfoxide (DMSO) as a control, and aliquots were drawn for scoring Kar9p polarity. Samples also were taken during the time course to monitor budding and progression of the spindle pathway (n = 300 cells). (B) Aliquots from the time course were analyzed for Kar9p distribution in short spindles after treatment for 15 min (nLatB = 268 cells; nDMSO = 204 cells) and 30 min (nLatB = 262 cells; nDMSO = 191 cells). More than 90% of cells exhibited Kar9p label at both poles after 30-min treatment with LatB. (C) Representative fields of cells treated with DMSO or LatB for 30 min from the experiment depicted in A. Treatment with LatB induced both spindle misalignment and symmetric localization of Kar9p (arrowheads). Bar, 2 μm.