Figure 2
Mifepristone adversely affects HEI-193 cells in culture. (a) Metabolic viability of HEI-193 cells decreases with increasing concentration of mifepristone after 72 h in culture. Individual data points represent metabolic activity as percentage of vehicle-treated control for seven independent experiments, performed in replicates of 3–5 wells per condition (vehicle-treated cells versus 35 μM mifepristone-treated cells, p = 0.006; **p < 0.01). (b) Phase object confluence remains constant among cells treated for 72 hours with 35 μM mifepristone, while vehicle-treated cells exhibit normal proliferation. Error bars in the mifepristone-treated condition are smaller than the size of the symbol. (c) 10X phase contrast images of vehicle-treated cells (top row) and cells treated with 35 μM mifepristone (bottom row) 6 h and 72 h post-treatment. (d–f) The percentage of BrdU+ HEI-193 cells significantly declines after 72 h treatment with mifepristone (p = 0.0007; ***p < 0.001): (d) 25X epifluorescence image of cells treated with 0.1% DMSO vehicle, with BrdU in red and Hoechst stain in blue; (e) Cells treated with 35 μM mifepristone, where the white box encloses a single BrdU+ cell; (f) Quantification of five replicate experiments. (g) Activation of fluorescent caspase 3/7 detected via live-cell imaging over 100 h of mifepristone treatment reveals a slight but not significant elevation in caspase 3/7 activity between mifepristone-treated cells (70 μM and 35 μM) and vehicle-treated control cells; positive control is staurosporine (0.5 μM) and drug is applied 48 h after plating. Note that after 96 h in culture, normally proliferating vehicle-treated cells become overly confluent and begin to apoptose. (h–i) Phalloidin staining of mifepristone-treated cells reveals crumpled f-actin morphology after 72 h: (h) Cells treated with 0.1% DMSO vehicle; (i) Cells treated with 35 μM mifepristone, where rhodamine phalloidin is red and Hoechst stain is blue. Center values in histograms are means; error bars are s.e.m.
