Fig. 5. Pro-inflammatory cytokines induce the expression of mesenchymal-like genes and diverse cellular phenotypes in a patient-derived PFA cell model.

a Experimental design. The PFA primary cell line EPD-210FHTC was cultured in the presence or absence of TGF-β1 and/or TNF-α for 5 days. The experiment was performed in three biological replicates. b Image of the EPD-210FHTC cells after the 5-day culture, for each of the treatments. TGF-β1 is required for the cells to acquire a mesenchymal phenotype, whereas treatment with TNF-α potentiates the formation of 3D colonies. c Average change in the gene expression level of MLC-specific markers with respect to the no-treatment control after 5 days of treatment. Gene expression levels were profiled by RT-qPCR. Both TGF-β1 and TNF-α induce the upregulation of MLC markers, but the relative expression levels of FN1 and CHI3L1 strongly depend on the particular treatment. Error bars indicate 90% confidence intervals. d Cell migration assay. An example of the same gap at day 0 and day 5 is shown for each condition. Treatment with TGF-β1 leads to a substantial increase in the invasive potential of the cells. e Average fraction of the area invaded by the cells after 5 days in the cell migration assay across four biological replicates (two-sided t-test; *p value $\le 0.05$, **p value $\le 0.01$). Error bars indicate 90% confidence intervals. f Cell proliferation assay by EdU incorporation. The average proportion of EdU⁺ cells after 2 days of treatment is shown for each of the experimental conditions. The cell proportions in each condition were compared to the cell proportion in the no-treatment control across three biological replicates using a two-sided t-test. Treatment with TGF-β1 and/or TNF-α leads to a small reduction in cell proliferation (**p value $\le 0.01$, ***p value $\le 0.001$). Error bars indicate 90% confidence intervals. Source data and numeric p values are provided in the Source Data file.