Additional file 5: Figure S5. of Multi-walled carbon nanotubes directly induce epithelial-mesenchymal transition in human bronchial epithelial cells via the TGF-β-mediated Akt/GSK-3β/SNAIL-1 signalling pathway

Representative microscope images of BEAS-2B cells (A-B) and relative expression of epithelial and mesenchymal markers (C). A-B) BEAS-2B cells were incubated for 96 h in either the absence (0 μg/ml, CTRL) or presence of [Fe3+] (released by FeCl3) similar to the one potentially released at the highest concentration of MWCNT used, Fe-depleted MWCNTg (44 μg/ml) and MWCNTg (44 μg/ml). After the incubation, the cells were rinsed with PBS and observed by OM (A) or fixed with paraformaldehyde and fluorescently labelled as described in Methods (B). A) Representative images are shown (10×; scale bar = 50 μm). B) Actin filaments were visualized in red, nuclei in blue. Representative images are shown (63×; scale bar = 10 μm). C). Relative expression of epithelial and mesenchymal markers were checked by WB. Relative expression of E-cadherin, β-catenin, α-SMA and vimentin proteins in BEAS-2B cells incubated in either the absence (0 μg/ml, CTRL, lane 1) or presence of [Fe3+] (released by FeCl3, lane 2), Fe-depleted MWCNTg (44 μg/ml, lane 3) and MWCNTg (44 μg/ml, lane 4). GAPDH was used as loading control for cytosolic extracts. Each figure is representative of three experiments giving similar results. (TIF 1558 kb