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Vps34 is required for the interaction of Sec7 and Rsp5 in vivo. Lysates prepared from VPS34 (LHY5440) or vps34Δ (LHY5508) cells expressing Sec7-GFP, or VPS34 cells expressing untagged Sec7 (LHY1850), were subjected to immunoprecipitation (IP) with GFP antibodies. Total lysates (0.7%, anti-GFP blot and 0.07%, anti-Rsp5 blot) and precipitated proteins were resolved by SDS-PAGE and analyzed by immunoblotting (IB) with GFP and Rsp5 antibodies.
