SIRT1 mRNA target binding measured by RABS using a trans-scaffold. (A) Observed base-pairing pattern of mSIRT1:miR-34a duplex as determined by NMR spectroscopy (Baronti et al. 2020). (B) Averaged reactivity values of three independent replicates for the unbound scaffold containing the mSIRT1 sequence (gray), RNA:RNA (red, +RNA) and RNA:RNA-RBP complexes (blue, +RNA-RBP), with one standard deviation indicated by shaded regions. (C) Reactivity values after normalization over buffer nucleotides of the main loop. Values >1 and <0 are set to 1 and 0, respectively. (D) Heatmaps of the same data presented in B. (E) ΔReactivity values in which the unbound scaffold reactivity is subtracted from the bound reactivity at each nucleotide. Nucleotides in the mSIRT1 target sequence interacting with the miR-34a seed region are indicated by green underlining and letters.
