Fig 7.

Effects of Lcn-2-conditioned medium on RL95-2 cell migration. RL95-2 cells (8 × 10⁴) were stimulated with 10 μM Lcn-2 for varying lengths of time, and then culture supernatants were collected from conditioned medium. Cytokine antibodies (anti-IL-8 or anti-IL-6) were added to clarify the effect of these cytokines on 48hrCM-induced cell migration. Cell migration was measured using the transwell assay in the absence or presence of conditioned medium. (A) After incubation for 24 h, RL95-2 cells were stained with 0.5% crystal violet, and cells were counted in ten random fields under a microscope at 20× magnification. (B) OD₅₇₀ values of cells on the lower surface of the membrane extracted with 33% acetic acid after anti-IL-8 neutralization. These data are representative of three independent experiments. The cell number of RL95-2 incubation under 48hrCM for 24h is as a control experiment. Values are the mean ± SEM. **, P < 0.01; ***, P < 0.001.