Fig. 4: CCL20 binds to integrin α5β1 to activate lung fibroblasts.

a List of the 10 proteins that interact with CCL20 identified by MS analyses. b Volcano plot of enriched proteins that interact with CCL20 (n = 3 technical replicates per group). Significantly altered proteins are shown in red (false discovery rate (FDR) < 0.05, log₂FC > 1) and blue (FDR < 0.05, log₂FC < −1). c Identification of integrin α5 and integrin β1 from CCL20 immunoprecipitates in fibroblasts by MS analysis. d Surface plasmon resonance (SPR) analysis of the kinetic interaction between CCL20 and integrin α5β1. CoIP analysis of the interaction between CCL20 and integrin α5 (e) or integrin β1 (f) in HEK293T cells. IF images showing the colocalization of CCL20 with integrin α5 (g) or integrin β1 (h) in MRC-5 cells. Scale bars, 10 μm. i Expression of Col1a1, Col3a1, Fibronectin and Acta2 in primary fibroblasts treated with or without ATN-161 in the presence of CCL20. j Immunoblots showing COL1 and α-SMA levels in primary fibroblasts treated with or without ATN-161 in the presence of CCL20. k Migratory ability of primary fibroblasts treated with or without ATN-161 in the presence of CCL20. Scale bars, 100 μm. l Expression of ACTA2 in MRC-5 cells treated with or without ATN-161 in the presence of CCL20. m Immunoblots showing the levels of COL1 and α-SMA in MRC-5 cells treated with or without ATN-161 in the presence of CCL20. n Migratory ability of MRC-5 cells treated with or without ATN-161 in the presence of CCL20. Scale bars, 100 μm. The data are presented as the means ± SEM. For (i–n), n   =   3 technical replicates per group. The data in i–n were analyzed by one-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001. Source data are provided as a Source data file.