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. 2015 Apr;135(4):998–1007.e6. doi: 10.1016/j.jaci.2014.08.030

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© 2014 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

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Fig 5 — Reptin and Pontin interact with AID in vivo. A, A schematic representation of the immune-isolation protocol for AID-interacting proteins. B, AID and co-purifying proteins were isolated from Flag-AID-transfected BOSC23 cell extracts by immunoprecipitation with anti-Flag antibodies and then analysed by Western blotting with the indicated antibodies. C, c-myc-tagged Reptin (Reptin_c-myc) and Pontin (Pontin_c-myc) were immunoprecipitated with anti-c-myc antibody from lysates of BOSC23 cells expressing Reptin_c-myc, Pontin_c-myc, and AID and then analyzed by Western blotting with the indicated antibodies. Anti-Clathrin and “beads-only” (‘/’) immunoprecipitations were used as negative controls. Reptin_c-myc and Pontin_c-myc were always co-transfected because these proteins stabilize each other⁴¹ (Fig E4). D, Endogenous Reptin interacts with AID in switching splenocytes. Anti-Flag immunoprecipitates from lysates of IL-4- and LPS-stimulated AID^−/− (−/−) and AID^Flag/Flag (F/F) splenocytes were analyzed with an antibody against endogenous Reptin.