Fig. 6

USP37 knockdown inhibits stemness, cell invasion and EMT via Hedgehog signaling pathway in breast cancer. a, b MCF-7 cells were incubated with 0.5 μM purmorphamine for 24 and 48 h. a Hedgehog pathway constituents were examined via western blotting. GAPDH was examined as a loading control. **P < 0.01,***P < 0.001. b Immunofluorescence staining images of MCF-7 cells showed the expression of USP37 and Hedgehog pathway constituents. c Protein levels of USP37, Smo, Gli-1, ALDH1, OCT4, E-cadherin, N-cadherin, Snail1 as detected by western blotting after the NC siRNA group or the USP37 siRNA#2 group was treated with 0.5 μM purmorphamine for 48 h. GAPDH was examined as a loading control. **P < 0.01,***P < 0.001. d Cell invasion capacity of the NC siRNA group or the USP37 siRNA#2 group treated with 0.5 μM purmorphamine (Scale bar: 200 μm). e Spheroid formation capacity of MCF-7-ShScramble or MCF-7-shUSP37#2 cells treated with 0.5 μM purmorphamine (original magnification, 4×). f Immunofluorescence staining of E-cadherin and N-cadherin after the NC siRNA group or the USP37 siRNA#2 group treated with 0.5 μM purmorphamine for 48 h. (Scale bar: 50 μm). *P < 0.05, **P < 0.01