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. 2020 Nov 30;15(11):e0242725. doi: 10.1371/journal.pone.0242725

Fig 9. PUF60 uses UHM to associate with SF1 and cooperatively interact with AdML3’ splice site region pre-mRNA.

Fig 9

(A) Representative isotherms from ITC experiments to detect the association between SF1 1–260 and either PUF60 RRMs+UHM or PUF60 RRMs. (Left panel) Titration of SF1 1–260 into PUF60 RRMs+UHM. The best fit of the binding stoichiometry (n) was one. All titrations were measured at 30°C. (Right panel) Titration of SF1 1–260 into PUF60 RRMs; (B) Reporting the thermal dynamic parameters measured in (A). The mean parameters and standard deviations of two experiments are reported. The standard state (°) for these experiments is defined as 1 M concentrations of each protein at 30°C. The dissociation constant (Kd) and stoichiometry of binding (n) are derived from curve fitting based on datasets subtracted with the reference titration of SF1 heat of dilution. a Calculated using the equation ΔG° = –RT ln(Kd-1). b Calculated using the equation ΔG° = ΔH°-TΔS°. (C) EMSA study of the interplay of SF1 1–260 and PUF60 RRMs+UHM with AdML3’ RNA. The bright-dark colors of the gel image are digitally reversed for clarity in presentation.