Extended Data Fig. 4: Normal lymph node HEV morphology and numbers upon Grem1+ FRC ablation.
From: Gremlin 1+ fibroblastic niche maintains dendritic cell homeostasis in lymphoid tissues
a, Representative plots with relative proportion (top) and absolute numbers (bottom) of CD45− EYFP+ stromal cells in Grem1-Cre-ERT2 Rosa26 EYFP (n = 5) and Rosa26 EYFP/iDTR (n = 5) mice upon DTxn mediated ablation. Plots are gated on CD45− stromal cells. b, Spleen weight (left) and cellularity (right) in Grem1-Cre-ERT2 Rosa26 EYFP (n = 12) and Rosa26 EYFP/iDTR (n = 14) mice upon DTxn mediated ablation. c, Immuno-fluorescence imaging of lymph node sections from Grem1-Cre-ERT2 Rosa26 EYFP (top) and Rosa26 EYFP/iDTR mice (bottom) upon DTxn mediated ablation, sections were stained for anti-LYVE-1(lymphatics, LECs) and anti-PNAd (HEVs). Scale bars, 150μm. d, Number of HEVs per lymph node as identified by PNAd staining as in (c), here comparing Grem1-Cre-ERT2 Rosa26 EYFP and Rosa26 EYFP/iDTR mice upon DTxn mediated ablation. e, Images of lymph node sections from Grem1-Cre-ERT2 Rosa26 EYFP (left) and Rosa26 eYFP/iDTR mice (right) upon DTxn mediated ablation, sections were stained for anti-PDPN (stromal cells) and anti-PNAd (HEVs). Scale bars, 25μm. f, Immuno-fluorescence imaging of lymph node sections from Grem1-Cre-ERT2 Rosa26 EYFP (left) and Rosa26 EYFP/iDTR mice (right) upon DTxn mediated ablation, sections were stained for anti-PDPN (stromal cells). Scale bars, 75μm. g, Relative proportion of ITGA2b + LEC in Grem1-Cre-ERT2 Rosa26 EYFP (n = 10) and Rosa26 EYFP/iDTR (n = 9) mice upon DTxn mediated ablation. Plots are gated on LEC: CD45− Ter119− CD31+ PDPN+. Data are representative of three independent experiments (a); pooled data from three independent experiments (b); Representative image and quantitation of ten lymph nodes from Grem1-Cre-ERT2 Rosa26 EYFP and fourteen lymph nodes from Rosa26 EYFP/iDTR mice from three independent experiments (c, d, f); representative image of seven lymph nodes from three independent experiments (e); pooled data from two independent experiments (g). ****P < 0.0001, **P < 0.01 (two-tailed unpaired Student’s t-test). Exact P-values in extended source data. Data are shown as mean ± SEM.
