1. FLC does not significantly inhibit AMR assembly and constriction and septin localization.
FLC does not significantly inhibit AMR assembly and constriction and septin localization. (A) To monitor constriction of the AMR, time-lapse microscopy was performed using a strain that expressed a component of the AMR, myosin heavy chain (Myo1), tagged with mCherry, and a nucleoporin (Nup107), tagged with GFP (to monitor the stage of mitosis; strain LC4). Cells from the control treatment (DMSO) had a constricted AMR. FLC-treated cells that formed multimeras also constricted the AMR, although the dynamics of the constriction differed significantly from those seen with untreated control cells. The arrow in the uppermost of the panels corresponding to the 70-min time point indicates a dark line that likely corresponds to a septum formed after the AMR had constricted. Bars, 5 µm. (B) Cells expressing septin Cdc10-mCherry (LK62) were pretreated with 24 μg/ml FLC for 6 h, and the localization of Cdc10-mCherry was analyzed by time-lapse microscopy while the cells were continually exposed to FLC (the times indicated represent the progression of the time lapse). Cdc10-mCherry formed a collar and a double ring between the mother and the first daughter (arrow 1) and formed another such ring when the second daughter was formed (arrow 2). (C and D) Cells deleted for Cdc3- or Cdc12-encoding genes (LK65, LK162) exhibited hypersensitivity to FLC (C), with a clear defect in cytokinesis at 24°C (D). Bars, 5 µm in panels A and B and 10 µm in panel D.
Sophie Altamirano et al. mSphere 2017; doi: /mSphere