Hedgehog–Gli1 pathway inhibition by genistein in vitro . (A) Real-time polymerase chain reaction analysis of human Smoothened (SMO) and Gli1 in MCF-7 cells. The cells were incubated with different concentrations of genistein for 48 hours. (B) Protein levels of SMO and Gli1 were measured by western blot analysis in MCF-7 cells and accompanied by a quantitative bar chart. As an internal control, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used for normalization. *P <0.05, #P <0.05, **P <0.01, Student’s t test.