Figure 6A–6D.

Scavenging ROS/RNS and stimulating autophagy enhanced the cytotoxicity of silibinin under a background of ERα activation. The cells were treated with 200 μmol/L of silibinin for 24 h in the presence or absence of 0.25 μmol/L PPT, 2.5 mmol/L NAC, 200 U/L SOD, 0.5 mmol/L L-NAME, 500 μmol/L 1400W or 10 nmol/L rapamycin. (A) Cell viabilities were measured by MTT assays. ^*P<0.05 vs silibinin and PPT treatment group. n=3. Mean±SD. (B) Cell death ratios were estimated by the trypan blue exclusion method. The quantitative cell death ratios are presented as means±SD. ^*P<0.05 vs the silibinin and PPT-treated group. The morphologic changes were observed by phase contrast microscopy (C), or fluorescence microscopy with AO staining (D), scale bar=20 μm.