Ectopic expression of Ndt80 does not profoundly accelerate the induction of middle sporulation gene transcription. RNA samples (10 μg) obtained at the indicated time points from wild-type NDT80-expressing cells (A) or wild-type cells that also express NDT80 under the regulation of an IME2 promoter, IME2-NDT80 (B), were probed for the middle sporulation gene transcripts SPS1 and SPS2. RNA samples (10 μg) from a homozygous ime2 mutant expressing NDT80 from its endogenous promoter, NDT80 (C), an IME2 promoter, IME2-NDT80 (D), or a CUP1 promoter, CUP1-NDT80 (E), were probed for the middle sporulation gene transcripts SPS1 and SPS2. The ACT1 transcript served as a loading control.