Extended Data Fig. 10: Characterization of Col3a1ΔN mice.
From: Matrix-producing neutrophils populate and shield the skin
a, Schematic representation of the Col3a1fl/fl mouse gene construct. Exons 2-3-4 are flanked by loxP sites which are recognized as targets of DNA cleavage by Cre recombinase. Primers 1 and 2 are used for the genotyping. Primers 3 and 4 are used for detection of depletion of gene construct after crossing with the Mrp8Cre driver line. b, Genotypes of Mrp8Cre Col3a1fl/fl (Col3a1ΔN) mice determined by PCR of genomic DNA using primers 1 and 2 that target the inserted loxP site (left). Deletion of the Col3a1 gene was confirmed by PCR of genomic DNA of lung neutrophils using primers 3 and 4 (right). Controls are CreNEG floxed neutrophils. DNA from lung fibroblasts was used as control for driver specificity. Blots are from 3 experiments. c, Representative 3D reconstructions of MRP14+ neutrophils and PDPN+ fibroblasts stained for Col3a1, with Col3a1+ cells indicated with asterisks (left). Percentage of Col3a1+ neutrophils and fibroblasts extracted from lungs of Cre-negative controls and Mrp8Cre; Col3a1fl/fl (Col3a1ΔN) mice (right). Data are presented as mean values ± s.e.m. and from 4 Cre-negative mice (455 neutrophils and 106 fibroblasts) and 5 Col3a1ΔN mice (505 neutrophils and 142 fibroblasts). d, Representative images and quantification of COL3+ matrix rings around day 3 skin wounds in the same control and Col3a1ΔN mice. Right, quantification of COL3+ and laminin+ areas (from Fig. 4h) is shown in the box and whisker plot. The number of replicates are displayed in the figure. P-values determined by two-tailed Student’s t-test. Box plots show median ± interquartile; whiskers show the range from minimum to maximum.
