Fig. 4: LYN deletion disturbed the CAF-like phenotype and associated polarization in stromal cells by diminishing inflammatory features and enhancing typical myofibroblastic functions.

a Heatmap of a curated list of genes associated with CAF phenotype. Fold changes (log₂-FC) from “omics”-layers are depicted. Red and blue color represent log₂-FC of genes in LYN^WT and LYN^KO clones, respectively. Individual layers, where a gene was not detected, are gray. Genes were grouped by their cellular compartment and asterisks indicate “omics”-layers where genes were significantly differentially expressed (specified in Methods). b Transcriptional levels of CAF markers according to Biffi et al.²⁴ in different LYN^WT and LYN^KO cells (i: HS-5, ii: NKtert, iii/iv: two imCAF lines) determined by qRT-PCR. (HS-5: 3 SCC per genotype; NKtert: 1 LYN^WT SCC vs. 2 LYN^KO SCC; imCAF#1: 2 polyclones per genotype, imCAF#2: 1 LYN^WT vs. 2 LYN^KO polyclones). c Immunoblot of LYN^WT or LYN^KO imCAF#1 and imCAF#2 for selected CAF markers. (i) and (ii) were prepared from the same lysates. d Immunoblot of LYN^WT or LYN^KO HS-5 cells (3 clones per genotype, untreated or stimulated with 10 ng/ml TGFβ for 24 h) for selected CAF markers. e (i) Bright field (left) and immunofluorescence microscopy (right) for phalloidin and Hoechst showing morphology of LYN^KO cells under normal culture conditions. (ii) Quantification of FSC and SSC of HS-5 LYN^WT and LYN^KO (mean ± SEM, 3 clones per genotype). f Migration of HS-5 cells in a scratch assay. The scratch area relative to the maximal value was measured every 20 min by live cell microscopy for HS-5 LYN^WT (1 clone) and LYN^KO (3 clones) in independent triplicates (mean ± SD) (i). Area Under the Curve (mean ± SD; Mann-Whitney test p = 0.0044) (ii). g LYN^WT and LYN^KO HS-5 cells were treated for 24 h with kinase inhibitors (1 µM dasatinib, 5 µM bosutinib, 5 µM saracatinib), mRNA levels of PDGFRB and FAP were assessed by qRT-PCR (2 SCC per genotype). In all qRT-PCR gene expression was normalized to PPIA. Depicted are means (± SEM where applicable) from the indicated number of clones per genotype. See also Fig. S3. Source data are provided as a Source Data file.