Fig. 6.

Effect of IP6K1 depletion on tumorigenic potential of cancer cells. (A) Anchorage-independent growth of cells seeded in soft agar was determined after 3 weeks by staining with crystal violet to visualize and count the number of colonies. Representative images of crystal violet stained colonies are shown. (B) Bar graph shows the number of colonies per dish in IP6K1 depleted HeLa and HCT116 cells normalized to their respective NT controls. Data are mean ± SEM from three independent experiments and were analyzed by a one-sample t-test. (C) Colonies formed in soft agar (A) were imaged at higher magnification as described in the Methods section. (D, E) Bar graphs showing the colony sizes obtained after growth of HeLa (D) and HCT116 (E) cells for 3 weeks in soft agar (n = 66 and 67 for HeLa NT control and shIP6K1–4 respectively; n = 63 and 52 for HCT116 NT control and shIP6K1–1 respectively). Data (mean ± SEM) are representative of two independent experiments and were analyzed using the non-parametric two-tailed Mann-Whitney test. (F) Subcutaneous xenografts of HCT116 NT (top row) and shIP6K1–1 (bottom row) cells, isolated 4 weeks after injection of cells into either flank of the same mouse. (G) Symbol and line plot displaying the weights of tumors derived from HCT116 NT and shIP6K1–1 cells. Symbols indicate the tumor weight scatter, and lines represent the pair-wise analysis of tumors isolated from either flank of the same mouse (n = 8). Data was analyzed using a two-tailed paired Student's t-test. **P ≤ 0.01; *P ≤ 0.05; ns, not significant, P ≥ 0.05.