Fig. 4.

H. diminuta extract (HdE) suppresses WKYMVm and KC-induced chemotaxis in murine bone marrow neutrophil. Freshly isolated neutrophils (2 × 10⁵ NØs) were placed in the outside wells of an ultrapure agarose gel and chemotaxis towards HdE or WKYMVm (1 μM) assessed (a upper panels are representative images showing extensive migration of neutrophils out of the well (demarcated by the dashed line) in response to WKYMVm but not HdE). b shows that NØs pretreated with HdE (100 μg/mL, 30 min) had reduced chemotaxis to WKYMVm. Using a transwell (8 μm pore size) migration system revealed that HdE inhibited NØ chemotaxis to WKYMVm (c) and KC (20 ng/mL; d), but not leukotriene B₄ (LTB₄, 100 nM; e). Treatment of NØs with HdE at the time of application of WYKMVm to the culture well (co-treatment) also suppressed chemotaxis (f; data are mean ± SD; each datum point is the average of 2 NØ replicates from an individual mouse; * and ^# p < 0.05 compared to control [con] or the appropriate chemotactic stimulus only, respectively, by one-way analysis of variance with Tukey's post-test).