Figure 2.

MDSC adhesion to endothelial cells after ENO1-treatment. (A) Bone marrow-generated MDSC were labeled with fluorescein-conjugated anti-CD11b Ab, and untreated or treated with anti-ENO1 mAb before seeding on TNF-α pre-activated endothelial cells for 1 h. Adherent cells were fixed and stained with crystal violet and counted in 10 fields/each condition. Graphs represent the mean ± SEM of two independent experiments in which 6 × 10⁴ and 3 × 10⁴ CD11b⁺ cells were seeded, respectively. ***p values < 0.0001, which significantly discriminate the ctrl- from ENO1-MDSC. (B) Representative pictures of CD11b⁺ cells untreated or treated with anti-ENO1 mAb in fluorescence (green; upper panels) and of the monolayer of endothelial cells in bright field (lower panels) at 10x magnification. (C) Adhesion to extracellular matrix components was assessed by seeding 1 × 10⁵ cells/well of ctrl- and ENO1-MDSC and CF-PAC-1, as a positive control, in duplicate on a 24-well pre-coated plate. After 90 min, adherent cells were washed and stained. OD was read at 570 nm. Bars represent mean ± SEM.