Figure 7. Western blot analysis of DNA damage responses to mre11A470T and ctf18▵ Mutations.

[Top]: Protein extracts were isolated from wild-type, mre11A470T, ctf18▵ and mre11A470T ctf18▵ cultures grown in 5 ml YPD cultures at 30°C or 15°C. Extracts were subjected to electrophoresis on a 10% SDS-PAGE gel prior to Western analysis using goat primary polyclonal anti-Rad53 antibody as described in Materials and Methods. Phosphorylated forms (arrows on right) migrate more slowly than the un-phosphorylated species. [Bottom]: Protein extracts were isolated from cold resistant mre11A470T ctf18Δ survivors and grown in 5 ml YPD at 15°C before shifting to 30°C. Extracts were subjected to Western analysis for Rad53 expression as described above.