Figure 6.

Treatment of B-cells with Nint impairs B-cell–driven fibroblast migration and activation. (A and B) Images of quiescent IPF fibroblasts stained with Wright-Giemsa stain migrating toward BCM, CPG-BCM, and Glu-BCM. Some BCMs were prepared by preincubating B-cells for 1 hour with 1 μM Nint or 100 μM Pirf before stimulation with CpG or β-glucan (CpG-BCM/Nint, CpG-BCM/Pirf, Gluc-BCM/Nint, and Gluc-BCM/Pirf), as indicated. Controls include regular medium (Med) and medium containing CpG, Gluc, Pirf, and Nint (Med/CpG, Med/Gluc, Med/Pirf and Med/Nint, respectively) and are shown in Figure E2. Nint was also added to the BCM only when the Transwell assay is set up to control for any Nint carryover (CpG-BCM/Nint+ and Gluc-BCM/Nint+). Each membrane was imaged and divided into four fields, migrated cells were counted in each field, and means are plotted as migrated cells per field. (C) Cell lysate of IPF fibroblasts stimulated with different BCMs (as indicated) and immunoblotted for FN1, αSMA, PAI1, or β-actin, as indicated. BCM was obtained as previously indicated (A and B). TGFβ was used as positive control. Data are representative of three separate experiments. ***P < 0.001.