Fig 4.

Quantification of Lcn-2 in RL95-2-conditioned medium. RL95-2 cells were cultured with or without Lcn-2 for the indicated times. Levels of Lcn-2 in conditioned media were determined by 12% SDS-PAGE and Western blotting with anti-Lcn-2 IgG. These data are representative of three independent experiments. (A) Lanes 1-3, conditioned media from RL95-2 cells cultured in serum-free medium for 24, 36 and 48 h, respectively; lanes 4-6, conditioned medium from RL95-2 cells cultured with serum-free medium supplemented with 10 μM Lcn-2 for 24, 36 and 48 h, respectively; lane 7, 20ng of purified Lcn-2; lane 8, cell-free control consisting of an equivalent amount of Lcn-2-supplemented medium incubated for 48 h at 37 ºC without cells. For lane 7, Lcn-2 was loaded on the gel. (B) mRNA levels of two Lcn-2 receptors, LRP-2 and Lcn-2R, were measured by RT-PCR analysis.