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Equilibrium dissociation constants measured by titrating substrate RNA with H/ACA RNPs. Sub-U (A) and Sub-Ψ (B), each at a 10 nM concentration, were incubated with WT-, D85A-, DEL7- and R154Q-RNP and their ΔGar1 counterparts of increasing concentrations for 30 min at 27°C. The fractions of bound substrates obtained from FCS analysis were fit to Equation (5) to derive the apparent dissociation constant Kd.
