FIG. 2.

Simultaneous overexpression of rho4⁺ and eng1⁺ or agn1⁺ causes dramatic cell lysis. (A) Wild-type cells transformed with plasmids pREP3Xeng1⁺ and pREP4X, pREP3X and pREP4Xrho4⁺, or pREP4Xrho4⁺ and pREP3Xeng1⁺ were grown at 32°C in EMM with 15 μM thiamine (nmt promoter off, top panel) or in EMM without thiamine (nmt promoter on, bottom panel) to induce rho4⁺ and eng1⁺ overexpression. (B) Images of wild-type cells transformed with plasmids pREP3X and pREP4X, pREP3Xrho4⁺ and pREP4X, pREP3X and pREP42Xagn1⁺, and pREP3Xrho4⁺ and pREP42Xagn1⁺. Cells were grown at 32°C in EMM without thiamine for 22 h and stained with methylene blue. Lysed cells appear dark. Bar, 10 μm.