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. 2004 May;24(10):4151–4165. doi: 10.1128/MCB.24.10.4151-4165.2004

FIG. 4.

FIG. 4.

Cell cycle- and DNA damage-dependent Sae2 phosphorylation in the absence of Mec1 and/or Tel1. Strains expressing SAE2-HA3 from the SAE2 promoter were as follows: wild type (YLL1103), tel1Δ (DMP3807/2D), mec1Δ sml1Δ (DMP3806/2A), and mec1Δ tel1Δ sml1Δ (DMP3916/5B). (A and B) Cell cultures growing logarithmically in YEPD were arrested in G1 with α-factor and released from the pheromone block at time zero in YEPD, or were UV-irradiated (40 J/m2) prior to the release in YEPD. Samples of untreated and UV-treated cell cultures were withdrawn at the indicated times after α-factor release in order to analyze the DNA content by fluorescence-activated cell sorting in nonirradiated (A, top) and UV-irradiated (A, bottom) cell cultures and to analyze Sae2 phosphorylation by Western blot analysis with anti-HA antibodies of extracts from nonirradiated (B, top) and UV-irradiated (B, bottom) cell cultures. (C) Cell cultures were synchronized with α-factor and transferred in YEPD containing α factor after UV irradiation (40 J/m2) (+UV + α factor). Time zero (αf) corresponds to cell samples taken immediately before UV treatment. Protein extracts from samples withdrawn at the indicated times were treated as described for panel B. exp, exponentially growing cells.