Figure 3.

Effect of 4-HC on cell proliferation and survival. A. B16-F10 cells were treated with 500 μM 4-HC (triangles) or 0.75% ethanol (control cells; squares) and viable cells were quantified every 12 h using the neutral red accumulation assay. Experiments performed in presence of 10% FBS (solid lines) showed that cell proliferation was unaffected by 4-HC (P > 0.05, ANOVA). When cells were treated in serum-free medium (dashed lines), the initial cell number was unaltered neither by time nor by 4-HC. Values are mean ± SE obtained from 3 independent experiments. B. Clonogenic assay were performed by seeding 4-HC-treated cells at a density of 20 cells/cm². After incubation for 8 d, the generated colonies were stained and counted. There was no difference in the colony number produced by 4-HC-treated cells or control cells. Values in the graph are mean ± SE obtained from 3 independent experiments. The upper picture shows examples of the results.