Figure 6

Cabozantinib overcomes gemcitabine resistance. (a) BxPc-3-GEM cells, which are a BxPc-3-derived and gemcitabine-resistant subclone, were left untreated or were treated with 10 μ M cabozantinib (XL), 50 nM gemcitabine (GEM) or both together (GEM+XL). Seventy-two hours later morphology was photographed at × 100 magnification. (b) At the same time point after treatment viability and apoptosis were analyzed as described in Figure 5a, and cell death was evaluated by examining granularity of cells by FACS-analysis. Specific death was calculated according to the formula given in Figure 2c. (c) Seventy-two hours after treatment, the expression of c-Met, phosphorylated Met (p-Met), cIAP2, Survivin and SOX2 were analyzed by western blot. (d) Likewise, seventy-two hours after treatment, cells were cytospinned to glass slides and the presence of cells positive for cleaved caspase-3 or Ki67 were analyzed by immunohistochemistry to detect apoptosis and viability, respectively. Photographs of positive cells (red and dark brown) are shown. The number of positive cells was quantified in 10 vision fields under × 400 magnification and the means±S.D. are shown