FIG. 5.

Benomyl treatment triggers changes in meiotic gene expression. (A) Northern blot analysis of IME2, RPL3, and SNR6 RNA levels. Wild-type cells (A1972) induced to undergo meiosis at room temperature in medium containing 0.4% DMSO (mock) (left) or 120 μg/ml benomyl (right). (B) The same strain as is shown in panel A was allowed to progress through meiosis for 4 h. After 4 h (black arrow), cells were resuspended in medium containing 0.4% DMSO (mock,) (left) or 120 μg/ml benomyl (right). Northern blot analysis of IME2, CLB5, CDC28, RPL3, and SNR6 RNA levels was carried out; rRNA was used as a loading control. rRNA levels were determined by staining with ethidium bromide; all other RNAs were detected by autoradiography. (C) tub2-150 cells (A5779) were pregrown at 30°C in medium containing 50 μg/ml benomyl and then sporulated at room temperature in medium containing 120 μg/ml benomyl. After 4 h, cells were resuspended in fresh medium containing 0.4% DMSO (mock) (top) or 120 μg/ml benomyl (bottom). Total RNA samples were analyzed by Northern blotting. Mock and benomyl-treated samples were run in the same gel but are separated in these figures for clarity. Total DNA content was analyzed by flow cytometry.