Figure 2.
The Involvement of Cyclin D1 in the Growth-Inhibitory Effect of LCAHA
(A) HCT116 p53wt (left panel) or HCT116 p53−/− (right panel) cells were treated with DMSO (marked with D), or 5 or 20 μM tested compounds for 48 hr, followed by western blot analysis. The graphs present densitometry analysis of cyclin D1 expression and show mean ± SEM from three independent experiments. Statistical significance was evaluated using one-way ANOVA with Tukey's post hoc test: *p < 0.05, **p < 0.01, ***p < 0.001. See also Figure S3.
(B) For the MTT assay the cells were seeded at low confluence and treated with tested compounds for 6 days. The graphs show mean values ± SD from three independent experiments.
(C) Analysis of cyclin D1 and p21 expression in the cells treated with DMSO or 10 μM LCAHA for 48 hr. The graph presents densitometry analysis of cyclin D1 expression and shows mean ± SEM from three independent experiments. For the statistics, a t test was performed: *p < 0.05, **p < 0.01.
(D) Colony formation assay was performed on MCF-7 or SAOS-2 cells treated with DMSO or 5 μM LCAHA for 5 days. The photographs are representative of three experiments.
(E) The numbers and sizes of the colonies formed in the colony formation assay (D) were measured using ImageJ software. Based on these data, the surviving fraction and relative mean colony size was calculated for the LCAHA-treated cells. The graphs show mean ± SE values from three experiments. For the statistics, a t test was performed: **p < 0.01; n.s., not significant.