Figure 4

Mifepristone treatment induces senescence in LNCaP cells. LNCaP cells were seeded at 50,000 cells per well and treated with mifepristone (MF) for 72 h followed by vehicle (VEH) for 5 days; or vehicle, 5% or 10% charcoal-stripped (CS)-FBS-containing media for 8 days. SA-β-galactosidase staining was performed as surrogate marker of senescent cells. Relative senescence was quantified as the number of cells with blue cytoplasm per field and expressed as the percent of total number of cells per field (A, B). Nine fields per well were quantified and completed in triplicate for each treatment group. All treatment groups were corrected against the background provided by the negative control, WI-38 cells that do not undergo senescence (data not shown). *** p < 0.0001 vs. VEH (one-way ANOVA followed by Tukey’s post-test). Scale bar = 50 μm.