FIG. 1.

DNA replication in HU-treated glc7 mutants. (A) Serial dilutions (1:10) of exponentially growing cultures of cells with the indicated genotypes were spotted onto YPD plates with or without MMS, HU, and phleomycin (phleo) and incubated at 25°C for 3 days. wt, wild type. (B to D) Exponentially growing wild-type, glc7-129, glc7-132, and glc7-T152K cells were arrested in G₁ with α-factor (αf) and released from the pheromone block in YPD or YPD containing 0.03 M HU. Aliquots of each culture were harvested at the indicated times after α-factor release to determine DNA content by fluorescence-activated cell sorting (FACS) analysis (B) and to detect Rad53 (C) and Ddc2 (D) by Western blot analysis with anti-Rad53 and anti-HA antibodies, respectively. exp, exponentially growing cells.