Extended Data Fig. 7: Characterization of TgfbrΔN mice.
From: Matrix-producing neutrophils populate and shield the skin
a, Blood counts in control and TgfbrΔN mice showing normal values for mice lacking neutrophil-specific TGFβ signalling. Data from 10 mice per group. b, Characterization of granulopoiesis in TgfbrΔN mice showing segmented, banded and metamyelocytic stages as percentages defined by imaging of Giemsa stains of bone-marrow samples. c, Differential expression of genes associated with blood, bone marrow (BM), spleen and lung signatures as defined in (3), as well as genes associated with the matrix signature (ECM) as defined in Fig. 1a. Comparisons are for neutrophils extracted from blood and lungs of wild-type control and TgfbrΔN mice. Data are from 3 mice per group. d, Representative reconstruction of neutrophils from the different tissues (top), and frequency of Col3a1+ neutrophils (bottom) extracted from tissues of wild-type control (n = 3) and TgfbrΔN mice (n = 3). e, Col3a1 expression in neutrophils imaged directly in the skin of wild-type control (822 cells from 5 images, 2 mice) and TgfbrΔN mice (1419 cells from 9 images, 3 mice). Representative images (left), showing neutrophils detected using Mrp14 which was used to normalize Col3a1 expression across samples (normalized intensity, middle dot plot). Using relative Col3a1+ levels >2,000 arbitrary units we stratified cells in tissues as Col3a1lo/hi and calculated the per cent of Col3a1hi cells in each group, where bars show median values. f, Representative images of Col3a1 expression inside and outside neutrophils imaged in the steady-state skin of Ly6GCreNEG control and Ly6G Cre+; Tgfbrfl/fl mice, showing neutrophils detected using Mrp14 (green), Col3a1 inside neutrophils (red) and Col3a1 outside neutrophils (magenta). g, Quantification of Col3a1 protein inside Tgfbr2fl/fl (143 cells, 2 mice) and Ly6GCre; Tgfbr2fl/fl neutrophils (275 cells, 3 mice); bars show median values. Right, box plots show extracellular Col3a1 protein signal in the same images from Tgfbr2fl/fl (143 cells from 11 images, 2 mice) and Ly6GCre; Tgfbr2fl/fl mice (275 cells from 17 images, 3 mice). P-values determined by two-tailed Student’s t-test. All controls here are Cre-negative Tgfbrflox littermates. Box plots show median ± interquartile; whiskers show the range from minimum to maximum.
