Fig. 3

The effect of IL-33 on the pro-fibrotic activity of fibroblasts. Non-IPF (n = 2) and IPF (n = 2) HLFs were stimulated with 2 ng/ml of TGFβ or 10 ng/ml of IL-33 for 4, 8 and 24 h and ACTA2 (A), COL1A1 (B), IL6 (C) and CXCL8 (D) gene expression measured by qPCR. For each donor, all data was normalised to the unstimulated media alone control at 4 h. Non-IPF and IPF donors are shown in black and grey respectively. Bars indicate median values and each data point represents a single donor with statistical analysis performed by Wilcoxon signed-rank test. Whole cell lysates from resting (rest.) M1 and M2 MDMs, activated (act.) M1 (IFNγ stimulated) and M2 (IL-4 stimulated) MDMs, HPAECs stimulated with 5 ng/ml of TNFα, 5 ng/ml of TGFβ and 0.1 ng/ml of IL-1β (TTI) and a single non-IPF HLF donor were separated by SDS-PAGE (20 μg protein/lane) and expression of ST2 and GAPDH measured by western blot (E). Whole cell lysates from HUVECs, non-IPF (n = 3) and IPF (n = 4) HLFs were separated by SDS-PAGE (20 μg protein/lane) and the expression of ST2 and GAPDH determined by western blot (F). Representative cropped western blots are shown