FIG. 3.
ScaANBS1 temporal expression during S-phase recovery is dependent on UvsBATR. (A) Western blot of total protein extracts from A. nidulans wild-type and scaA::pyr4 strains. The wild-type strain was grown for 30 min in the presence or absence of 25 μM CPT. (B) Real-time RT-PCR specific for A. nidulans nimA gene. Conidia from the wild-type strain (GR5) were grown in a reciprocal shaker at 37°C for 16 h in YG+UU medium. Mycelia were aseptically transferred to fresh YG+UU medium plus 20 mM hydroxyurea (HU) for 6 h (at 37°C). A control without HU was also made (0*). Mycelia were extensively washed with sterile YG medium and then transferred to fresh YG+UU medium at 37°C in a reciprocal shaker. Samples were collected every 10 min for 120 min and then the RNAs were extracted. (C) Western blot of total protein extracts from the A. nidulans wild-type strain (GR5). The protein samples were obtained from mycelia grown as described for panel B (lane 1, time zero; lanes 2 to 13, 10 to 120 min). The membrane was separately labeled with the anti-ScaA and anti-NpkA antibodies. (D) Densitometric analyses of the Western blots are shown in panel C. The results are ratios of the intensities of ScaA signals to the intensities of NpkA signals. (E) The ΔuvsB strain (AAH14) was grown as described for panel B, and a Western blot of the total protein samples was separately labeled with the anti-ScaA and anti-NpkA antibodies (lane 1, time zero; lanes 2 to 13, 10 to 120 min; lane 14, 18 h).