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. Author manuscript; available in PMC: 2009 Oct 30.
Published in final edited form as: Nature. 2009 Mar 8;458(7242):1201–1204. doi: 10.1038/nature07844

Figure 3. Stable expression of ZFP809 in 293A cells leads to reconstitution of the PBS silencing complex in vivo.

Figure 3

(A) Nuclear extracts from clonal cell lines control Clone 1 (empty vector control), Clone 5 (cell line stably expressing FLAG-tagged ZFP809(1-353) or Clone 9 (cell line generated with same construct as Clone 5 but which does not express ZFP809) probed by western blot with antisera for FLAG (upper panel), TRIM28 (middle panel), or beta-actin (lower panel). (B) Nuclear extracts prepared either from PCC4 EC cells or cell lines control Clone 1, Clone 9, or Clone 5 used in EMSA reactions with radiolabeled probes corresponding to PBSPro (PRO) or PBSB2 (B2). In final three lanes the PBS silencing complex is supershifted by the addition of 2 µg of either anti-TRIM288, anti-FLAG, or a control anti-HA antibody. (C) ZFP809(1-353) and TRIM28 co-immunoprecipitate in DNase treated Clone 5 whole cell extracts in the presence of ethidium bromide. Control immunoprecipitations were performed with of HA antisera. Immunoprecipitated ZFP809(1-353) and Co-immunoprecipitated TRIM28 was detected by western blot.