Fig. 1

Identification of miRNAs critical for an EMT and cell migration. a Expression profiling of miRNAs during a TGFβ-induced EMT in NMuMG/E9 cells. NMuMG/E9 cells were treated with TGFβ for the time points indicated, and total RNA was isolated for miRNA sequencing. The heat map summarizes the hierarchical clustering of differentially expressed miRNAs (log2FC(±2); FDR <0.05) compared to epithelial, untreated cells during an EMT according to the indicated colour scale. b Identification of miRNAs controlling EMT. NMuMG/E9 cells were transfected with miRNA mimics for individual miRNAs and analysed for mesenchymal characteristics in the absence and presence of TGFβ for 4 days. Formation of focal adhesions (green), actin cytoskeleton reorganization to actin stress fibres (red), fibronectin deposition (yellow) and nuclei (blue) were visualized by high-content fluorescence screening microscopy. c Identification of miRNAs regulating mesenchymal tumour cell migration. Mesenchymal, migratory Py2T cells (>20 days TGFβ) were transfected with different miRNA mimics, plated in 96-well Boyden chamber migration inserts within a FCS gradient and in parallel on a 96-well input plate. Cell nuclei were imaged using a fluorescence screening microscope and quantified. Migrated cells were normalized to the total cell number on the input plate. d Scheme depicting the workflow and the number of (a) differentially expressed miRNAs during an EMT, (b) the number of identified miRNAs functionally contributing to a TGFβ-induced EMT and (c) the number and names of the miRNAs functionally contributing to mesenchymal tumour cell migration