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. 2005 Nov;16(11):5227–5235. doi: 10.1091/mbc.E05-05-0405

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Copyright © 2005, The American Society for Cell Biology

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Figure 3. — Effect of mec1-85 mutation on interaction among Mec1, Ddc2, and Rfa1. (A) Effect of mec1-85 mutation on the Mec1–Ddc2 complex formation. Extracts prepared from cells expressing tagged or untagged Mec1 and Ddc2 proteins were immunoprecipitated with anti-HA antibodies. Immunoprecipitate (IP) and extract were subjected to immunoblotting analysis with anti-HA or anti-myc antibodies. Strains used here were MEC1-HA (KSC1635), MEC1-HA DDC2-myc (KSC1642), mec1-85-HA DDC2-myc (KSC1643), and DDC2-myc (KSC1644). (B) Effect of mec1-85 mutation on the interaction between Mec1 and Rfa1. Extracts were prepared from cells expressing tagged or untagged Mec1 and Rfa1, and immunoprecipitated with anti-FLAG antibodies. Immunoprecipitate (IP) and extract were then subjected to immunoblotting analysis with anti-FLAG or anti-HA antibodies. Strains used were RFA1-FLAG (KSC1655), MEC1-HA RFA1-FLAG (KSC1656), mec1-85-HA RFA1-FLAG (KSC1657), MEC1-HA (KSC1658), and mec1-85-HA (KSC1659). (C) Effect of mec1-85 mutation on the interaction between Ddc2 and Rfa1. Extracts were prepared from cells expressing tagged or untagged Ddc2 and Rfa1 and immunoprecipitated with anti-FLAG antibodies. IP and extract were then subjected to immunoblotting analysis with anti-FLAG or anti-HA antibodies. Strains used were RFA1-FLAG (KSC1655), DDC2-HA RFA1-FLAG (KSC1652), mec1Δ DDC2-HA RFA1-FLAG (KSC1653), mec1-85 DDC2-HA RFA1-FLAG (KSC1654), and DDC2-HA (KSC1651).