Fig. 3

Differential regulation of epithelial and mesenchymal markers by DUSP6. a Immunoblot analysis of proteins DUSP6, p-ERK, ERK, vimentin, N-Cadherin, E-Cadherin, fibronectin in long-term cultures U87MG treated with different stimuli (serum, starvation, EGF 100 ng/mL, cisplatin (CDDP) 5 µg/mL) for 5 h (left lane) or 24 h (right lane). b Immunoblot analysis of mesenchymal and epithelial markers in U87MG adenovirus-expressing DUSP6 compared to TRK (empty vector). The cells were treated with increasing doses of cisplatin (CDDP) at different concentrations (respectively 0, 2, 5, 10 μg/mL) and protein expression assayed by western blot on total lysates at 5 h. c Invasion assay. Bar graph reports the mean of total number of invaded cells per field compared to control (MOCK i.e. U87MG not infected). U87MG cells were transduced with adenovirus-expressing DUSP6 or control vector (TRK i.e. U87MG infected with empty vector) and seeded on matrigel-coated trans-well inserts. The experiment was performed for 48 h and the invading cells were counted (10× objective) in nine randomly chosen microscopic fields per trans-well. Data are presented as mean ± S.D. from two independent experiments performed in duplicate