Figure 3

Cell proliferation and cyclin D1 expression was lower in 7E-treated ML-1 cells. (a) ML-1 cells were seeded and incubated with IL-20 (200 ng/ml), 7E (2 μg/ml), or IL-20 (200 ng/ml) plus 7E (2 μg/ml) for 48–72 h. Cell proliferation was determined using an MTT assay. Medium alone was used as an untreated control. *P < 0.05 versus the untreated controls. Values are means ± SD. Data are representative of three independent experiments. (b) The effects of 7E on cell cycle distribution were measured using flow cytometry. Data are representative of three independent experiments. (c) ML-1 cells were treated with IL-20 (200 ng/ml), 7E (2 μg/ml), or IL-20 (200 ng/ml) plus 7E (2 μg/ml) for 6 h, and the expression levels of cyclin D1 were analyzed using qRT-PCR with specific primers. Quantification analysis of mRNA was normalized with GAPDH. *P < 0.05 versus the untreated controls. Values are means ± SD. Data are representative of three independent experiments. (d) ML-1 cells were treated with IL-20 (200 ng/ml), 7E (2 μg/ml), or IL-20 (200 ng/ml) plus 7E (2 μg/ml) for 24 h. Cyclin D1 and p21^WAF1 protein levels were assessed using immunoblotting. β-actin was a loading control. Cropped blots were displayed.