Inflammation enhances B-cell sensitivity to CXCL12. (A) CXCL12 was measured in bone marrow extracellular fluid from 1 femur and tibia. Data are pooled from 3 experiments. (B-C) Twelve hours after challenging mice with IFA or cecal puncture, total bone marrow pooled from 3 mice was plated in a transwell to measure migration toward CXCL12. Data are representative of 3 independent experiments. (D) Chemokinetic potential was examined by adding CXCL12 in equal concentrations to both the transwell top and bottom chambers. Graph is representative of 2 independent experiments. (E) Chemotaxis assays for CXCL13 and CCL21 as compared with CXCL12. Each point represents the mean fold change in chemotaxis for an independent experiment. (F) IFA-treated mice were injected IP with 1 mg/kg AMD3100 1 hour prior to analysis. Data are pooled from 2 experiments. (G-H) B-cell expression level of CXCR4 was determined by flow cytometry measurement of median fluorescence intensity. (G) Error bars indicate standard error of the mean (SEM) and graph is a representative experiment, where n = 5. (H) Each point and connected line is the median fluorescence intensity (MFI) for a single experiment (n = 3-5 per experiment). (I) Cells were cultured for 1 hour in media containing 2.5 mM methyl-β-cyclodextran. Shown is a representative experiment.