Fig. 5: Bufalin blocks MMPs, MAPK and EMT signaling pathways in HepG2 cells.

a Bufalin-induced G2/M phase arrest in HepG2 cells. The cell cycle distribution was determined by flow cytometric analysis and cell cycle distribution was quantified. b Bufalin decreased the protein expression of CDK1, cyclinB1 and increased the protein expression of P53. Cells were exposed to bufalin (10 nM) for indicated time points and protein levels were detected by western blot. c Bufalin inhibited the formation of F-actin cytoskeleton in HepG2 cells. Arrows indicated F-actin (red) stained with rhodamine-phalloidin (Scale bars =10 μm). d Bufalin decreased the protein expression of MMP2 and MMP9. Cells were treated with or without bufalin (10 nM) for indicated time points. e Bufalin inhibited ERK/JNK/P38 MAPK signaling cascades. Non-phosphorylations/phosphorylations of ERK, JNK, and p38 were detected by western blot. f Bufalin inhibited EMT signaling pathway. Cells were treated with or without bufalin (10 nM) for 24 h. The levels of Vimentin, TCF/ZEB1, Snail, N-cadherin, E-cadherin, Claudin-1 were evaluated by western blot. Data are representative of three independent experiments. **P < 0.01 vs. control group; ns, not significant by ANOVA with Student’s t-test.