Fig. 4: DNASE1L3 expression in the GALT subepithelial region.
From: Double-negative B cells and DNASE1L3 colocalise with microbiota in gut-associated lymphoid tissue
a Imaging Mass Cytometry (IMC)-derived images illustrating spatial structure of an appendix follicle, including subepithelial dome (SED) (green dashed line) and germinal centre (GC) (grey dashed line), shown through expression of CD3 (T cells, brown), Ki67 (proliferation, cyan) and E-cadherin (epithelium, orange). b Same field as in (a) showing staining of CD68 (red) and E-cadherin (orange), as well as gene expression of DNASE1L3 (green) and ITGAX (encoding for CD11c, blue) through RNAscope. c Enlarged profiles of SED and GC regions from (b) showing individual and merged marker expression. d Quantification of distance to epithelium for cells segmented on CD68 and ITGAX classified as either DNASE1L3+ or DNASE1L3-. P = 0.0024, n = 14 ROIs. e Quantification of DNASE1L3 expression on cells segmented on CD68 and ITGAX classified as either GC or non-GC P = 6.25899 × 10−6, n = 10 ROIs. f IMC-derived images of ileum follicle illustrating CD3 (brown) and E-cadherin (orange) protein localisation alongside DNASE1L3 (green) and ITGAX (blue) gene expression, with GC highlighted (grey dashed line). g Visualisation of DNASE1L3 (green), NOX2 (blue) and lysozyme (red) distribution within the same image as in (a). h Enlargement of SED region depicted in (g). i Quantification of the average expression of NOX2 and lysozyme in DNASE1L3+ cells and classified as residing in either the SED or in non-SED regions P = 0.0107 and 7.5789 × 10−5 respectively, n = 14 ROIs. j Confocal microscopy image showing DAPI (blue), CD20 (cyan, pseudocolor), ITGAX (green) and CD11c (red) expression in an appendix follicle. ITGAX+CD20+ cells assessed using RNAscope are highlighted with cyan arrows. CD11c+ITGAX+CD20- cells are highlighted with red arrows. Statistics between groups were assessed by two-tailed t test. *P < 0.05, **P < 0.01 and ****P < 0.0001. Scalebars = 100 μm. Findings shown in (a, b, c, f, g) are representiative of 16 ROI. Data shown in (j) are representative of 6 ROI. Source data are provided as a Source Data file.
