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. Author manuscript; available in PMC: 2013 Mar 1.
Published in final edited form as: J Immunol. 2012 Jan 27;188(5):2380–2386. doi: 10.4049/jimmunol.1102454

Figure 1. HMGB1 Chemotaxis of MEFs and primary mature macrophages requires their production of CXCL12.

Figure 1

Migration assays primary WT MΦ (A) and immortalized WT MEFs (B) were performed in 48 well microchemotaxis chambers as previously described (16) in response HMGB1 (50 ng/ml), PDGF (10 ng/ml) (MEF migration positive control) or C5a (2 nM) (MΦ migration positive control) for 3 hrs. Neutralizing monoclonal antibody against CXCL12 (K15C) was added to the cells where indicated at 30 μg/ml, which effectively blocks CXCL12 activity (27). Migration results with 30/μg/ml of a mouse IgG2A isotype control antibody are also shown in Panel A. Bars are distances migrated through filter pores (WT MΦ in A) or net cell movement per High Power Field (HPF) after subtracting basal migration in serum free media (WT MEFs in B). Experiments were repeated 3–5 times each in duplicate. All error bars are standard error of the mean. Mean values of WT MΦ basal migration distances towards serum free media controls were 35 ± 2.6 and 35 ± 3.3 μm with and without K15C respectively and 31 ± 1.6 μm for mouse IgG2a controls. Mean values of WT MEF basal migration towards serum free media controls were 26 ± 1.1 and 22 ± 1.4 cells per HPF with and without K15C respectively. P values were determined by two (Panel A) or one (Panel B) way ANOVA with Tukey's post test (***p = <0.001).