Figure 2.
Concentration response assay for 2D monolayers of ovarian cancer cells treated with paclitaxel
(A) Representative images of OVCAR8 2D monolayer culture in the multichambered microwell chip 72 h after the addition of paclitaxel (upper panel) or DMSO control (lower panel) at the concentrations indicated. Cells were stained with the viability dye TMRM (magenta) and the apoptosis indicator caspase-3/7 (green).
(B) Specific viability after 72 h for OVCAR8 cells treated with paclitaxel at increasing concentration fitted with the Hill equation to evaluate IC50. Dotted line represents the predicted curve with R2 = 0.99, LogIC50 = −7.77, and H value = 1.34.
(C) Time course of specific viability of OVCAR8 cells exposed to different paclitaxel concentrations for 72 h.
(D) Time course of the specific apoptotic index (A.U., arbitrary unit).
(E) Confocal images of OVCAR8 cells cultured for 8 h with 1 × 10−6 M paclitaxel (upper panel) or without drug (lower panels). 3D rendering was performed in Imaris using conventional settings (left panel) showing microtubules (α-tubulin, gray) and nuclei (DAPI, cyan) or surface rendering with filament analysis (right panel) to visualize the poles of the mitotic spindles (dots in magenta).
(F) Fraction of multipolar OVCAR8 cells after 8 h cell culture in increasing concentrations of paclitaxel (n = 36 per concentration). Statistical method was one-way ANOVA followed by Dunnett’s post hoc test (∗∗∗∗p ≤ 0.0001).
(G) Example images of tubulin and nuclei staining of OVCAR8 cells cultured for 8 h in different concentrations of paclitaxel.