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. 2003 Aug;14(8):3280–3291. doi: 10.1091/mbc.E03-03-0154

Figure 4.

Figure 4.

Effect of actin depolymerization on Hsl1p and Hsl7p. (A) Cells of strain DLY5000 (HSL1-myc HSL7-HA) were grown to exponential phase and treated with 100 μM Lat-A for 1 h at 30°C and then fixed and double stained to visualize the septin Cdc11p and either Hsl1p-myc (top) or Hsl7p-HA (bottom). Bar, 10 μm. (B) G1 cells of strain DLY5000 were isolated by centrifugal elutriation and inoculated into fresh medium at 30°C. Lat-A or dimethyl sulfoxide (control) was added at 45 min (before bud emergence) and the incubation was continued for a further 2 h, and cells were processed to detect Hsl1p-myc (top) or Hsl7p-HA (bottom) by Western blot. (C) Cells of strain DLY5000 were grown to exponential phase at 30°C, treated with 100 μM Lat-B, and processed to detect Hsl1p-myc (top) or Hsl7p-HA (bottom) by Western blot. At later times, the population was skewed toward unbudded cells because some division took place but bud emergence was blocked. (D) Cells of strain DLY4339 (cdc12-6 HSL7-HA) were grown to exponential phase at 23°C, shifted to 37°C for the indicated times, and processed to detect Hsl7p-HA by Western blot.