Figure 4.

MiRs-143 and -145 overexpression alters cell junctions enhancing cell motility. (a-d) miR-143, miR-145 overexpressing and empty vector control NMuMG cells were analyzed by: (a) fluorescence with phalloidin-FITC (scale bar, 20 μm); (b) cell proliferation assay (mean cell counts±S.E.M, n=3); (c) western blot for EMT and junction proteins; (d) scratch assay in a confluent cell monolayer. Images (left panel) take advantage of the endogenous RFP encoded by the miRNA expression constructs (scale bar, 100 μm); histograms (right panel) show the % of wound closure relative to the empty vector control (mean±S.E.M, n=3). (e,f) NMuMG cells were silenced for ZO1 with three independent shRNA constructs, or infected with a scrambled control (e), and analyzed by scratch assay assessed as above (f). P-values: *P<0.05; ***P<0.001