Figure 3: UNR facilitates the interaction of MLE with roX2 in vitro.

(a) Schematic representation of roX2 and derivatives used for RNA pulldown. (b) Titration of MLE in the presence or absence of UNR. MS2-tagged SL67 RNA was immobilized on amylose beads and incubated with MLE and UNR. Complexes were eluted as described in the Methods section. The effect of UNR on MLE binding to rox2 is visible at low MLE concentrations. (c) Correlation between UNR and MLE binding. MS2-tagged SL67 and SL2345 were used for RNA pulldown in the presence of MLE and/or UNR, and complexes were eluted. Bound MLE and UNR were quantified relative to the amount of eluted RNA, and normalized to the value obtained for SL67. Quantifications correspond to the average of two independent experiments. (d) UNR promotes the binding of MLE to full-length roX2. UNR and MLE were allowed to bind to full-length, ΔSL6 and ΔSL7 roX2 RNAs, and the RNPs were eluted. The percentage of bound UNR and MLE was quantified relative to the amount of eluted RNA. Values were normalized to MLE and UNR binding to full-length roX2. s.d.’s were determined from three independent experiments. Full-size blots corresponding to this Figure are shown in Supplementary Fig. 6.