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. 2014 Nov 12;9(11):e112762. doi: 10.1371/journal.pone.0112762

Figure 1. Flag-6xHis-tagged HBZ represses Tax transactivation and can be purified.

Figure 1

(A) The functional activity of Flag-6xHis-HBZ was determined by measuring its ability to repress Tax trans-activation of a LTR1-luc reporter in 293T cells. Cells were transfected with S-tag Tax (200 ng), LTR-1 luciferase reporter (100 ng) renilla-TK (20 ng) and a titration of HBZ constructs (100 and 500 ng.) Values represent mean measurements relative to Tax expression alone. Error bars represent standard deviation. Western blots were performed on 20 ug total cell lysate and probed with rabbit HBZ-specific antisera. Actin was used as a loading control. (B) Flag-6xHis-HBZ was purified from lysates of transfected 293T cells using agarose beads conjugated with anti-Flag antibody. Bands were resolved by SDS-PAGE and the gel was stained with GelCode Blue.