Figure 4

miR-200c-3p negatively regulates cell binding to fibronectin. (A) The cell binding to fibronectin coated on V-bottom wells was measured and compared between the HEK293T cells transduced with miR-200c-3p pmiRZip, miR-200c-3p mimic, or empty vector (control). The percentages of bound cells were determined as described in the Methods section, and the relative ratios to control vector-transduced cells are shown. (B, C) The cells were incubated with HUTS-4 (mAb that interacts with the activation-dependent epitope of β1 integrin) or mouse isotype control antibody (IgG2b). Bound mAbs were detected with allophycocyanin-conjugated secondary antibody and analyzed using flow cytometry. Representative FACS histograms from three independent experiments are shown (B). MFI values for HUTS-4 mAb were determined and relative levels of HUTS-4 binding to control vector-transduced cells were compared (C). (A, C) Data are expressed as the mean ± standard errors of the mean (SEM) relative to the control vector (Cont. vec.). All assays were performed in at least triplicates and the experiments were repeated three times. MFI, mean fluorescence intensity. ****P < 0.0001.