FIGURE 4.
CEACAM1 on NETs is important in migration of colon carcinoma cells. (A) Boyden chamber assays of HT-29 cells on NET monolayer isolated from human neutrophils stimulated with 500 nM PMA and treated with 200 μg/ml 5F4 mAb (5F4), 200 μg/ml isotype control of the 5F4 mAb (IgG), or 10 μM Sivelestat (NEi). Shown are the mean and mean SE of five fields per condition done in triplicate. *p < 0.05, NS compared with stimulated neutrophils. (B) Representative fluorescent images of the migrated CFSE-HT-29 cells of the four conditions presented in (A). Scale bar, 20 μm. (C) Boyden chamber assays of MC38-CC1L cells on PMA-stimulated (light gray bar) or non-stimulated (black bar) neutrophils from C57BL/6 (WT) or PMA-stimulated (white bar) or non-stimulated neutrophils (dark gray bar) from CC1 KO mice. Shown are the mean and mean SE of four fields per condition done in triplicate. *p < 0.05. (D) Representative bright field images of crystal violet stained MC38-CC1L migrated cells of the four conditions depicted in (C). Scale bar, 20 μm.