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. 2020 Sep 16;123(11):1644–1655. doi: 10.1038/s41416-020-01040-y

Fig. 3. Enhancement of metastatic ability and activation of epithelial-to-mesenchymal transition (EMT)-related signalling pathways in breast cancer cells after osteogenic induction.

Fig. 3

a Cell migration was measured by scratch-wound analysis at the indicated time. Scale bars = 500 µm. b The migration and invasion capacities were determined by transwell assay and quantified by counting the number of cells perforated crystal violet at the indicated time points after OC treatment. Scale bar = 200 µm. Western blotting (c) and qRT-PCR (d) of EMT-related genes in MDA-MB-468 and MCF-7 cells at various time points after OC induction. e Transforming growth factor (TGF)-β levels in MDA-MB-468 and MCF-7 cell supernatants were assessed by collecting and mixing supernatants at days 3 and 5 of OC induction. f The activation of the EMT-related Smad2/3 and MAPK (e.g., JNK, ERK and p38) pathway in MDA-MB-468 and MCF-7 cells was measured by western blotting at various time points upon OC treatment. β-actin was used as an internal control. *P < 0.05, **P < 0.01, determined by unpaired Student’s t test and one-way ANOVA with Bonferroni post-test correction. Data are shown as mean ± SD, and they represent three independent experiments with similar results.