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. Author manuscript; available in PMC: 2013 Mar 13.
Published in final edited form as: Mol Cell. 2010 May 14;38(3):439–451. doi: 10.1016/j.molcel.2010.04.012

Figure 1. Purification of HIV-1 Tat Complexes.

Figure 1

(A) HIV-1 Tat was purified from Dignam nuclear extracts prepared from Flag-HA epitope-tagged Tat (eTat)-expressing HeLa S3 cells (S3Tat) or nontransduced S3. eTat was sequentially purified on anti-Flag and anti-HA antibody-conjugated agarose beads. Proteins were resolved by SDS-PAGE and visualized by silver staining. The identity of eTat-associated proteins was determined by mass spectrometry (see also Figure S1 and Table S1).

(B) Flag IP from samples shown in (A) were resolved on SDS-PAGE, and the presence of eTat-associated proteins identified was confirmed by WB.

(C) 7SK RNA copurifies with eTat. Total RNA was extracted from samples used in (A). The presence of 7SKRNA was assessed by RNase protection assay using a full-length 7SK probe.