Figure 4
Decreased clonogenic and tumorigenic potential of mesenchymal DU 145 SKP2 KD cells is accompanied by a decrease of CD44+CD24− cancer stem cell subpopulation. (A) Skp2 and p27Kip1 expression in SKP2 KD cells were determined by western blot. Representative images are from three independent experiments. β-actin was used as a loading control. (B) Analysis of CD44 and CD24 markers in control and SKP2 KD cells using flow cytometry. Dot plots show representative results from three independent experiments. (C) Quantitation of results shown in (B). Data are presented as the mean ± SD of the percentage of CD44+CD24− cancer stem-like cells (n = 3). (D) The capability of spheroid formation and growth in anchorage-independent conditions in SKP2 KD cells and control cells. (E) Quantitation of results shown in (D). Data are shown as the mean ± SD of the number of spheroids per well. Results are from three independent repetitions. (F) The xenograft formation rate of SKP2 KD cells in SHO mice. Tumour formation rate was then calculated for control and SKP2 KD cells. A number of mice per group, n = 10. (G) ALDH1 activity in DU 145 mesenchymal cells with Skp2 knocked down. Data are presented as the mean ± SD of the percentage of cells with active ALDH1. Results are from three independent experiments.
