Figure 3. Bcr binds to UAP56.
(A) After over expressing Bcr and UAP56 in HeLa cells, total cell lysates (TL) were prepared and UAP56 and Bcr were immunoprecipitated using respective antibodies and Western blotting was performed with UAP56 antibody. (B) (Mammalian two-hybrid assay). HeLa cells were transfected with pBIND or pBIND-Bcr and pACT UAP56 full length or UAP56 fragments. Cells were harvested and luciferase assay performed. Full length UAP56 binds to Bcr as does UAP56 fragment 2 (amino acids 101-200). (**p<0.01). (C) HeLa cells were transfected with pBIND-Bcr and pACT UAP56 full length or UAP56 fragments as indicated (the pACT vector contains the herpes simplex virus VP16 activation domain). Bcr was immunoprecipitated with Bcr antibody and immunblot done with UAP56 antibody. Over expression of UAP56 fragment 2 [amino acids (aa) 101-200] blocked Bcr/UAP56 binding. Over expression of fragment 3 (aa 201-300) had no effect.