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Fig. 4. Analysis of the Rlc1-GFP rings in different strains. (A) Left panels: WT, myo3
, and myo3
chs2
cells were stained with Calcofluor and photographed under a conventional microscope. The position of Rlc1-GFP or Rlc1-GFP and the septum is shown. The arrows point to secondary rings that are slightly displaced from the septum. Right panels: different confocal images of a three-dimensional reconstruction of the Rlc1-GFP rings in WT, chs2
, myo3
and myo3
chs2
strains. The arrows point to deformed rings. (B-D) Time-lapse analysis of CAR contraction in different strains. Cells from WT (B; bar 5 µm), myo3
(C), and myo3
chs2
(D) strains were photographed at the indicated times (in minutes). The arrows and diamond indicate cells in which Rlc1-GFP can be visualized from the first stages of ring assembly. The asterisk and the spots mark cells in which the ring was already assembled. Bar, 10 µm. (E) A graphical representation of the average time for ring assembly or ring contraction and disassembly in different cells from the indicated strains.