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. 2006 Jul;80(14):6855–6864. doi: 10.1128/JVI.00383-06

FIG. 3.

FIG. 3.

The N terminus of Rep68 specifically binds ANP32B in vivo. (A) Coimmunoprecipitation experiments. Lysates from HEK293 cells transfected with the combinations of plasmids indicated at the top were immunoprecipitated (IP) with an anti-Flag antibody and Western blotted with an anti-HA antibody or immunoprecipitated with an anti-HA antibody and Western blotted with an anti-Flag antibody, as indicated. Immunoprecipitates were treated with MNase for 30 min at 37°C. Western blot assays of the input whole-cell lysate (WCE) are also shown as a measurement of the total input material. (B) Schematic representation of the Flag-tagged isoforms of Rep used in panel C. Residues 1 to 224 of Rep contain the DNA binding domain (DBD); Rep40 is the shortest naturally occurring isoform of Rep, encompassing residues 225 to 526, which still include an SF3 viral helicase domain. (C) Binding of Rep68 and Rep40 to ANP32B by immunoprecipitation. Lysates from HEK293 cells were transfected with the indicated combinations of plasmids to test whether Flag-Rep40 was able to interact with HA-ANP32B. Western blot assays of input whole-cell lysate (WCE) are also shown as a measurement of the input material.