Figure 4. C3aR is required for efficient chemotaxis.

(a) BMDCs (5×10⁴) from WT mice were suspended in media (RPMI-1640) and were placed on 48-well chemotaxis chambers. After 1 hr. of incubation with C3aR agonist (CAS 944997-60-8; 100ng/ml) or vehicle (RPMI-1640) transmigrating BMDCs were detected in stained membranes, visualized under microscope and reported as the average number of cells/field. **P<0.01 as determined by the student t test. (b) BMDCs (5×10⁴) from WT mice were placed on 48-well chemotaxis chambers. After 1 hr. of incubation with BAL fluid supernatant pooled from WT or C3^−/− mice exposed to 6 month of air or smoke, transmigrating BMDCs were detected in stained membranes and were visualized under microscope and reported as the average number of cells/field. ***p<0.001, as determined by one-way ANOVA test with Bonferroni's multiple comparison. (c) BMDC (5×10⁴) from WT and C3ar^−/− mice were placed on 48-well chemotaxis chambers. After 15-, 30- and 60 min of incubation with BAL fluid supernatant pooled from WT mice exposed to 6 month of air or smoke, transmigrating BMDCs were detected in stained membranes, visualized under microscope and reported as the average number of cells/field. ***p<0.001 as determined by one-way ANOVA test with Bonferroni's multiple comparison. Results are represented as mean±s.e.m, from 3 independent experiments (n=6-8).