Extended Data Fig. 10. LTβR-stimulation regulates lung repair and regeneration by modulating WNT/β-catenin-signalling.

a, Schematic representation of the experiment in which B6 mice were exposed to FA (n=5) or CS for 6m plus control Ig (n=5), LTβR-Ig fusion protein (80 μg i.p., weekly, n=5), LTβR-Ig fusion protein + beta-catenin/CBP inhibitor PRI-724 (0.6mg i.p., 2x weekly, n=6) or CHIR99021 (0.75mg i.p., weekly, n=5) from 4 to 6m, and analysed at 6m. b, mRNA expression levels of Ltb, Tnfsf14 (Light) and Ltbr relative to Hprt, determined by qPCR in whole lung from the mice described in (a) (FA n=5, CS plus control Ig n=5, LTβR-Ig n=5, LTβR-Ig + PRI-724 n=6 and CHIR99021 n=5 mice/group). c, Representative images of immunohistochemical analysis for CD3 positive T cells and B220 positive B cells (brown signal, haematoxylin counter stained, scale bar 100μm) in lung sections from the mice described in (a). d, mRNA expression levels of Axin2 relative to Hprt, determined by qPCR in whole lung from the mice described in (a) (FA n=5, CS plus control Ig n=5, LTβR-Ig n=5, LTβR-Ig + PRI-724 n=6 and CHIR99021 n=5 mice/group). e, Schematic representation of human lung organoid experiments. f, Representative images and quantification of lung organoids from primary human alveolar type 2 epithelial cells cultured for 14d +/− human LTβR-Ag (2 μg/ml) and LiCl (5mM), (scale bar 500μm, n=2 replicates from 2 separate donors). g, Schematic representation of the re-ignition of repair and regeneration pathways in AT2 lung cells following LTβR-Ig therapy in both young and aged exposed to chronic CS. Data shown mean ± SD (d), P values indicated, two-tailed Student’s t test (d) and one-way ANOVA multiple comparisons Bonferroni test (f).