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. 2019 Nov;25(11):1509–1521. doi: 10.1261/rna.072256.119

FIGURE 4.

FIGURE 4.

SF3A1-UBL binds to G-C rich stem–loop RNA. (A) All competitive EMSA reactions contained 500 nM GST-UBL and 10 nM WT Cy5-U1-SL4. Final concentrations of competitor RNAs for WT U1-SL4, Drosophila melanogaster (Dm), and two mutant RNAs, M10 and M10h, were 0.02, 0.03, 0.06, 0.13, 0.25, 0.5, and 1 µM. The dissociation constants of competitor RNAs (KC) that competed out Cy5-U1-SL4 are provided (n = 3). (B) Dose-response curves plotted from the fraction of Cy5-U1-SL4 bound versus the concentration of competitor RNA are shown. (C) Sensorgrams from SPR experiments for binding of GST-UBL to Dm, M10, and M10h RNAs. GST-UBL concentrations were 0.02, 0.03, 0.06, 0.13, 0.25, and 0.5 µM. Orange traces represent the global fit to the raw data by the Langmuir binding model, assuming a 1:1 stoichiometry. Binding kinetics and dissociation constant values are summarized below sensorgrams where binding was detected.