Endogenous miR-503 and VEGF production with various stimulations.

<p>HFL-1 cells were cultured in monolayer for 2 days, after which the medium was changed to DMEM containing 0.2% FCS, with or without IL-1ß and TNF-α (1 ng/ml), TGF-ß1 (1 ng/ml) or PGE₂ (1 x 10⁻⁷ M). (A) 1 day after stimulation, RNA was isolated and miR-503 expression was analyzed by real-time qPCR. MiR-503 expression was calculated as fold of 18s-rRNA expression. Vertical axis: level of miR-503 expression, expressed as fold of control medium alone. Horizontal axis: culture conditions. *<i>p</i> < 0.05 compared with the values of control medium alone. (B) HFL-1 cells cultured in monolayer were transfected with miR-503 inhibitor (Black bar) or control miRNA (White bar) transfection reagent, as described in Materials and Methods. 24hr after transfection, the medium was changed to DMEM containing 0.2% FCS, with or without TGF-ß1 (1 ng/ml), IL-1ß and TNF-α (1 ng/ml) or PGE₂ (1 x 10⁻⁷ M). 3 days after transfection, the culture medium was harvested and assayed for VEGF by ELISA. Vertical axis: VEGF release (pg per 10⁵ cells per 2 days). *<i>p</i> < 0.05, ***<i>p</i> < 0.001 compared with the values of control medium alone. The data represent the means ± SE of 3 separate experiments.