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. 1998 Jan 26;140(2):355–366. doi: 10.1083/jcb.140.2.355

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The assembly of the actin ring occurs downstream of Cdc15. RLY238 cells (cdc15-2 CYK1-myc) were grown at 37°C for 3 h. 5 ml of cells were fixed as the 0 time point sample. The rest of the cells were harvested and resuspended in 23°C YPD and grown at 23°C for 15 and 30 min before fixation or processing for immunoblot analysis. The fixed cells were double stained with anti-myc antibody and with rhodamine phalloidin. (A) Representative cells from the 0 time point that had a Cyk1 ring (top left). Representative cells from the 30-min time point that had a Cyk1 ring (bottom left). (B) The number of budded cells (mostly large-budded) that had a Cyk1 ring or an actin ring was counted and divided by the total number of budded cells counted (∼150) to yield the percentage. The number of unbudded cells were also counted and divided by the total number counted to yield the percentage. All cells that had an obvious actin ring had a Cyk1 ring. (C) Cells from each time point were processed for immunoblot analysis using anti-myc antibody. Bar, 10 μm.

The assembly of the actin ring occurs downstream of Cdc15. RLY238 cells (cdc15-2 CYK1-myc) were grown at 37°C for 3 h. 5 ml of cells were fixed as the 0 time point sample. The rest of the cells were harvested and resuspended in 23°C YPD and grown at 23°C for 15 and 30 min before fixation or processing for immunoblot analysis. The fixed cells were double stained with anti-myc antibody and with rhodamine phalloidin. (A) Representative cells from the 0 time point that had a Cyk1 ring (top left). Representative cells from the 30-min time point that had a Cyk1 ring (bottom left). (B) The number of budded cells (mostly large-budded) that had a Cyk1 ring or an actin ring was counted and divided by the total number of budded cells counted (∼150) to yield the percentage. The number of unbudded cells were also counted and divided by the total number counted to yield the percentage. All cells that had an obvious actin ring had a Cyk1 ring. (C) Cells from each time point were processed for immunoblot analysis using anti-myc antibody. Bar, 10 μm.