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. 2008 Feb 3;36(5):1599–1609. doi: 10.1093/nar/gkn010

Figure 2.

Figure 2.

tRNA-binding activity of recombinant RIC9. (A) Northwestern blots of purified recombinant RIC9 (100 pmol) were probed with the indicated 32P-labeled tRNAs (106 c.p.m./ml). (B) Gel-shift assay of refolded RIC9 (5 nM) incubated with indicated amounts of tRNATyr in 10 μl of buffer BB without added salt. (RNP = tRNA–RIC9 complex). (C) Scatchard plot of bound/free (b/f) against bound RNA concentration (b). Data from the gel-shift assay in panel (B). The slope of the best-fit curve = −1/Kd. (D) Effect of KCl on the binding of tRNATyr (1 nM) to 5 nM of recombinant RIC9 (upper panel) or RIC1 (lower panel). (E) Effect of heparin (10, 50 and 100 μg/ml) or E.coli tRNA (1, 5 and 10 nM) on binding of tRNATyr (1 nM) to RIC9 (5 nM).