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. 2014 Jun 13;462:1–13. doi: 10.1016/j.virol.2014.05.022

Fig. 3.

Fig. 3

MMLs stimulate the RNA binding activity of protein A. (A,B) MMLs stimulate the binding of protein A to RNA1(50–118) probe. Gel mobility shift assays were performed using increasing concentrations (wt/vol) of MMLs with 0.5 μM MBP-protA and 20 nM RNA1(50–118) probe. The concentrations of MMLs are indicated above each lane. The RNA binding activity of protein A in the absence of MMLs is used as the control (1-fold). The increase in the RNA binding activity of protein A at each point concentration of MMLs is graphed as the fold of control as shown in (B). Error bars represent S.D. values from at least three independent experiments. (C,D) MMLs stimulate protein A cooperatively binding to RNA1(50–118) at high protein concentrations. Gel mobility shift assays were performed using increasing molar concentrations of MBP-protA incubated with 0.2 μM RNA1(50–118) probe with the addition of 2 μg/μl MMLs, and then analyzed via 2% agarose gel to clearly separate migration of protein–RNA complexes. The Hill coefficients of the RNA binding of protein A in the presence of MMLs at low and high protein concentrations are indicated (D).