Fig. 2

Cirsiliol induces apoptosis in OS cells. A Annexin-V-FITC staining was used to determine the proportion of apoptosis occurring in U2OS and U2OS/MTX cells treated with DMSO (10 and 20 µM). B Statistical plots of early and late apoptosis in U2OS versus U2OS/MTX at different concentrations of cirsiliol for three replicate experiments. C Under the same concentration of cirsiliol treatment, a statistical plot of the difference in cell apoptosis between U2OS and U2OS/MTX cells was shown. D, E Western blot analysis of lysates from cirsiliol-treated U2OS or U2OS/MTX with BCL2, cleaved CASP3, cleaved CASP9, BAX and ACTB antibodies. Data are expressed mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001