Extended Data Fig. 9: Cellular activity of pre-miR-155-RiboTAC in MCF-10a cells overexpressing miR-155 and in HUVECs.
From: Programming inactive RNA-binding small molecules into bioactive degraders
a, Effect of pre-miR-155-RiboTAC on pre- and mature miR-155 in MCF-10a cells forced to express wild-type pre-miR-155, a binding site mutant of pre-miR-155 (A bulge to AU base pair), or a RNase L cleavage site mutant (pyrimidine-rich asymmetric loop mutated to base pairs) (n = 3 biological replicates). b, Forced expression of pre-miR-155, a mutant pre-miR-155 in which pre-miR-155-binder’s binding site has been mutated, or a mutant pre-miR-155 in which the RNase L cleavage site has been abolished enhances the migratory nature of MCF-10a cells, a model of healthy breast epithelium (n = 3 biological replicates). c, pre-miR-155-RiboTAC impairs the migration of MCF-10a cells forced to express WT pre-miR-155 but not those forced express the binding site or cleavage site mutants (n = 3 biological replicates). d, left: Effect of pre-miR-155-RiboTAC on mature miR-155 levels in HUVECs, as determined by RT-qPCR (n = 3 biological replicates). right: Effect of pre-miR-155-binder on the cleaving capacity of pre-miR-155-RiboTAC, as assessed by measuring pre-miR-155 levels by RT-qPCR (n = 4 biological replicates). e, Effect of pre-miR-155-binder on mature and pre-miR-155 levels in HUVECs, as determined by RT-qPCR (n = 3 biological replicates for mature miR-155; n = 4 biological replicates for pre-miR-155). f, Effect of control RiboTAC pre-miR-155-Ctr on mature and pre-miR-155 levels in HUVECs, as determined by RT-qPCR (n = 3 biological replicates). g, Effect of pre-miR-155-RiboTAC (100 nM) on the levels of mature miRNAs that harbour miR-155’s A bulge in their precursors in HUVECs, as determined by RT-qPCR (n = 3 biological replicates). h, Representative Western blot and quantification thereof of the effect of pre-miR-155-RiboTAC on pre-miR-155’s downstream target, Von-Hippel Lindau (VHL) (n = 3 biological replicates). i, Effect of pre-miR-155-RiboTAC on the angiogenic capacity of HUVECs (n = 2 biological replicates). All data are reported as the mean ± S.D. All p-values were calculated using a two-tailed Student’s t-test.
