Fig. 8: Human adhesion fibroblasts are heterogeneous and JUN dependent.

a Uniform manifold approximation and projection (UMAP) plots showing single-cell (sc)RNA-seq data from human adhesion fibroblasts FACS-isolated using an unbiased, lineage-negative sort strategy from three unique human specimens. Four unique clusters are identified. b UMAP plot showing representation of individual patient samples (arbitrarily numbered) across the cluster presented in 1. Human hashtag labels indicated at right, 2 hashtag antibodies were used for patient 33 as an internal control, as noted in the figure panel legend. c Pseudotime analysis of human abdominal adhesion fibroblast scRNA-seq data. Colors match cluster colors assigned in panel a. d Quantitation of qPCR analysis for JUN, STAT5, STAT3, and SPP1 of vehicle control versus JUN CRISPR Cas9-knockdown human abdominal adhesions fibroblasts. P-values noted in figure. n = 3 replicates per condition, datapoints represent average of technical replicates. e Quantitation of Ki67 expression using ICC of primary human abdominal adhesions fibroblasts treated with CRISPR Cas9 JUN knockdown compared with vehicle control. n = 4 biological replicates assessed per condition. f Quantitation of collagen type 1 expression using ICC of primary human abdominal adhesions fibroblasts treated with virally mediated JUN overexpression, vehicle control, or CRISPR Cas9 JUN knockdown. n = 3 biological replicates assessed per condition. g Schematic (based on published KEGG pathways) displays proposed JUN-relevant signaling pathways identified in this study. The left panel shows the acute phase response following tissue injury by which JUN is initially activated. The right panel shows the chronic profibrotic state that is established in adhesion fibroblasts. Blue circle highlights AP-1, red indicates the role of JUN inhibitor. Colors as labelled in the figure. ECM extracellular matrix. Data and error bars represent means ± SD. HPF high power field. A.U. arbitrary units. *P = 0.04, **P = 0.03, ***P = 0.03, unpaired two-tailed t-test. Source data are provided as a Source Data file.