Figure 2.

Effect of kaempferol and/or growth factors on DNA synthesis and apoptosis in HT-29 cells. (A, B) HT-29 cells were plated in 96-well plates, serum-starved and treated with 0 or 60 μmol/L kaempferol and/or growth factors [A: 10 nmol/L insulin-like growth factor-I (IGF-I); B: 20 μg/L heregulin (HRG)-β)] in the presence of [³H]thymidine. The cells were incubated for 3 hours and then harvested, and DNA synthesis was measured by incorporation of [³H]thymidine into DNA. (C, D) HT-29 cells were plated in 24-well plates, serum-starved, and treated with 0 or 60 μmol/L kaempferol and/or growth factors (C: 10 nmol/L IGF-I; D: 20 μg/L HRG-β). At 24 hours after treatment, the cells were trypsinized and loaded with Annexin V and 7-amino-actinomycin D (7-AAD). The percentages of early apoptotic cells (Annexin V⁺/7-AAD⁻) were quantified by flow cytometry. Each bar represents the mean±SEM (n=6). *P < 0.05 as compared with the untreated cells.