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Fig. 3. Identification and characterization of lad1-1 suppressors. (A) lad1-1 cells containing pUR19 (vector), pUR19 rho1 or pUR19gyp10 were isolated at 25°C and struck to YE plates at 36°C. Colonies were allowed to form for 3 days. (B) The pREP81-GFPgyp10 plasmid was transformed into wild-type cells (KGY246). After growth of transformants in the absence of thiamine for 16 hours, images of live cells were captured, and a composite of representative cells is shown. (C) Wild-type (KGY246), gyp10{Delta} (KGY2111), exo70{Delta} (MBY919) and gyp10{Delta} exo70{Delta} (KGY3945) cells were struck to plates for 3 days at the indicated temperatures. (D) gyp10{Delta} cells (KGY2011) were grown at 36°C for 6 hours, fixed, and stained with DAPI and Aniline Blue to visualize DNA and cell wall material, respectively. (E) Wild-type cells (KGY246) were transformed with the indicated plasmids and colonies recovered at 25°C. These were then grown in liquid medium at 36°C in the absence of thiamine for 18 hours. Cells were fixed with ethanol and stained with Aniline Blue and DAPI.