Fig. 2

Candidate miRNAs repress pathway activity and pathway-dependent growth. A. Stable isogenic Recombinant (SiR) MDA-MB-231 cells were transfected with miRNA mimics or treated with iCRT-14. 30 h later cells were stimulated with recombinant WNT3a. 18 h later, FLuc and RLuc activities were assayed. The effect of miRNAs and iCRT-14 are shown on normalized luciferase activity relative to respective negative controls. Significance was calculated by one-sample, two-tailed t-test on three independent SiR clones. P-values *** ≤ 0.0001, *** ≤ 0.001. B. MDA-MB-231, SUM-159, and HCC-1806 cells were transfected with miRNA mimics, 5 hours later medium was changed and, where marked, pathways were stimulated. 72 h post-transfection cells growth was evaluated by nuclei counts. The effect of miRNAs was compared to two negative miRNA mimics. Each experiment was repeated in biological triplicate, with six technical replicates each. Growth reduction is represented in red and growth induction in green. Corresponding full bar charts are shown in Supplementary Fig. 3 with statistics. C. MDA-MB-231, SUM-159, and HCC-1806 cells were treated with compounds or vehicle controls in combination with pathway stimulations, where marked. 72 h later cell growth was evaluated by nuclei counts. For every condition, the effect of treatments was compared to relative vehicle (DMSO for iCRT-14 and Capmatinib, PBS for Erlotinib; BSA as vehicle control for all stimulations). Being a relative growth, a single control bar is shown in plots. Each experiment was repeated in biological triplicate, with six technical replicates each. Significance was calculated by one-sample, two-tailed t-test. P-values ** ≤ 0.01, * ≤ 0.05. non-significant are not marked