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. 2012 Nov 29;8(11):e1003048. doi: 10.1371/journal.ppat.1003048

Figure 1. Purification and identification of LANA binding proteins.

Figure 1

(A) Outline of biochemical purification baits all of which pulled down Hsp90; (1.) wild-type, unmodified LANA; (2.) as used to derive partners listed in table 1; (3.) as used to derive partners listed in table 2. The abbreviations denote epitopes: F for DYKDDDDK, HA for YPYDVPDYA and EQEQE which is repeated within the LANA central domain. (B) Schematic procedure for purification of LANA complexes using bait 2. Samples were eluted from columns of Sepharose 6B and Heparin FF for purification separately, followed by immunoaffinity purification with mouse anti-Flag M2 affinity gel and immunoprecipitated using anti-LANA mab. (C) SDS-PAGE analysis was performed by 8–16% gradient gel and stained with colloidal blue. (D) Schematic procedure for purification of LANA mutant complexes using bait 3. (E) SDS-PAGE analysis was performed by 8–16% gradient gel and stained with colloidal blue. Co-IP, immunoprecipitation; M, molecular mass (in kDa).