Figure 7.

IL-6 and VEGFA participate in B-cell–mediated fibroblast migration and activation. (A) Images of quiescent IPF fibroblasts stained with Wright-Giemsa stain migrating toward different concentrations of recombinant IL-6 and IL-8 (A) or stimulated with different preparations of BCM, as indicated. Some fibroblasts were preincubated with 5 μM axitinib for 1 hour before the addition of the BCM, as indicated. (B) VEGFA secretion was measured by IB in the cell supernatant of NTC or stimulated cells with CpG and β-glucan for 24 hours as indicated. Some cells were pretreated with Nint and Pirf, as indicated. Nonspecific protein was used as loading control. Band intensities were quantified using ImageJ software version 1.48 (National Institutes of Health), and the relative intensities were calculated by normalizing the intensity of the target protein to that of the loading control. (C) Images of quiescent IPF fibroblasts stained with Wright-Giemsa stain migrating toward different concentrations of recombinant VEGFA or stimulated with different preparations of BCM, as indicated. (D) Cell lysates from IPF fibroblasts stimulated with different concentrations of IL-6 and VEGFA and immunoblotted for FN1, αSMA, PAI1, or β-actin, as indicated. Data are representative of three separate experiments. *P < 0.05, **P < 0.01, and ***P < 0.001. VEGFA = vascular endothelial growth factor A.