Fig. 3

miR-193b regulates apoptosis, migration, and stemness in TNBC cell lines. A and B – MDA-MB-231 cells were treated and 72 h later apoptosis was evaluated by Propodium Iodide positive nuclei. For each condition, the effect of treatments was quantified relative to respective controls. Unstimulated conditions represent BSA-containing media to the same final concentration present in stimulation conditions, where it was used as carrier protein. AThe effect of miR-193b was compared to two negative miRNA mimics. B The effect of inhibitors was compared to the respective vehicle controls. Each experiment was repeated in biological triplicate, with six technical replicates each. Significance was calculated by two-tailed t-tests (one-sample for chemical inhibitors, unpaired for miRNA OE). P-values *** ≤ 0.001, ** ≤ 0.01, * ≤ 0.05. non-significant are not marked. C. Serum-starved MDA-MB-231 cells overexpressing miR-193b or treated with iCRT-14 were seeded in the upper compartment of a transwell system, with serum in the lower chamber as chemoattractant. 20 h later, migration relative to controls was evaluated by nuclei counts. The effect of miR-193b was tested against miRNA mimic negative control #2 (top), and the effect of iCRT-14 against its vehicle, DMSO (bottom). The experiment was repeated in biological triplicate with three technical replicates each. Significance was calculated by one-sample, two-tailed t-test. P-values **** ≤ 0.0001. D and E – FACS analysis of CD24 and CD44 surface marker expression in MDA-MB-231 cells 96 h post-transfection with miR-193b or iCRT-14 treatment, compared to respective controls. The experiment was repeated in six biological replicates, with two technical replicates each. Significance was calculated by paired, two-tailed t-test. P-values indicated on each graph. P-values *** ≤ 0.001, ** ≤ 0.01, * ≤ 0.05. non-significant are not marked. D For each condition, the percentage of cells gated as CD44 positive and CD24 negative (stem-like population) is plotted. E For each condition, the percentage of cells gated as CD44 negative (left bars) or CD24 positive (right bars) are plotted