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. 2006 Jul 3;174(1):19–25. doi: 10.1083/jcb.200602049

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Copyright © 2006, The Rockefeller University Press

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Figure 4. — Pfa4 is required for palmitoylation of Chs3. (A) Lysates from cells expressing C-terminally 3xHA-FLAG epitope-tagged Chs3 from the GAL1 promoter were subjected to the acyl-biotin exchange reactions. Protein extracts were treated with (+) or without (−) hydroxylamine (NH₂OH). α-FLAG immunoprecipitates were blotted with α-biotin and α-HA to detect modified and total Chs3, respectively. (B) Cells expressing Chs3-GFP and Pfa4-3xHA were immunoprecipitated with α-HA antiserum and analyzed by Western blotting with α-HA and α-GFP mAbs. (C) Chs3 palmitoylation was assessed in wild-type, pfa4Δ, and chs7Δ cells by acyl exchange, as in A. (D) Coimmunoprecipitation of Chs3-GFP and Pfa4-3xHA in wild-type and chs7Δ cells as performed in B.