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. 2010 Oct 14;6(10):e1001152. doi: 10.1371/journal.ppat.1001152

Figure 4. The 5′ hairpin of human 7SK snRNA contains two HEXIM-binding sites.

Figure 4

A. Detection of HEXIM-binding sites by mobility shift assays. About 2 fmol of in vitro synthesized probe RNAs representing the entire (5′HP) or the distal (Dist) and proximal (Prox) parts of the 5′ hairpin of human 7SK were incubated with increasing amounts (fmol) of recombinant HEXIM1 and analyzed on a 5% native gel. Appropriate cold RNAs were used as specific competitors. B. In vitro interaction of mutant 7SK 5′ hairpin RNAs (5′HPdm, 5′HPpm, 5′HPdm+pm) with HEXIM1. Complexes were analyzed on a 4% gel. C. The minimal HEXIM1-binding elements of 7SK. The in vitro HEXIM-binding capacity of a distal (DHBS) and proximal (PHBS) fragment of the 7SK 5′ hairpin was tested by mobility shift assay on a 4% gel. Sequences derived from wild-type 7SK are boxed. D. Gelshift analysis of an artificial hairpin RNA (5′HPsyn) carrying the distal and proximal HEXIM-binding motifs of 7SK. Sequences originated from the human 7SK snRNA are boxed.