Fig. 6

The effect of VCAM-1 on BMDM migration. A Phase-contrast image showing the morphology of BMDM cultures. B, C Flow cytometry analysis of BMDMs. Live cells were gated on FSC-A and SSC-A to remove debris (B) and before plotting their CD11b and F4/80 expression (C). D Diagram of the migration assay using the Boyden Chamber system. BMDMs were added into a transwell insert with a porous membrane. The recombinant VCAM-1 protein (25 μg/ml) was added into the bottom chamber. The anti-VCAM-1 Ab (20 µg/ml) was added to the upper chamber. E Representative images of migrated macrophages on the bottom side of the inserts stained with DAPI from each treatment group. F Bar graph showing the number of migrated cells in different groups. *P < 0.05, n = 3. Mean ± SD. Two-way ANOVA followed by Turkey’s multiple comparisons test. G Experimental design to detect the effect of blocking VLA-4 using BIO5192 in VCAM-1-induced macrophage migration. H Representative images of migrated macrophages on the bottom side of the inserts stained with DAPI from each treatment group. I Bar graph showing the number of migrated cells in different groups. *P < 0.05, n = 3. Mean ± SD. Two-way ANOVA followed by Turkey’s multiple comparisons test