Extended Data Fig. 1: HFD-associated changes to intestinal villi and lacteals, and adipocytes and FALC in VAT.
a, Representative immunofluorescence images of intestinal villi. Cell nuclei (Hoechst, grey), CD31 + blood vessels (pink) and LYVE-1+ lacteals (green). Scale bars, 100 µm. b-e, Quantification of the width and length of the intestinal villi and lacteals from immunofluorescence images. f, Representative immunofluorescence images of VAT tissue stained for lipid droplets (green, Bodipy C16 FA). Scale bar, 50 µm. g, Quantification of adipocyte size (µm2). Mean ± s.e.m, n = 4, 5 mice in (b-c), n = 4, 4 mice in (d-e) with 3-4 intestinal villi analysed per mouse, n = 3, 3 mice in (g). Statistical differences, *p < 0.05 from two-tailed Mann-Whitney test. h, Immunofluorescent images of podoplanin + (green) lymphatic vessels in mesenteric adipose tissue from lean patient 2 and 3 and obese patient 2. Images from lean patient 1 and obese patient 2 are in Fig. 1c. (i-m) Representative immunofluorescence images of lymphatic vessels (LYVE-1 + , pink), blood vessels (CD31 + , cyan), CD11b + myeloid cells (grey), CD4 + T cells (yellow), B220 + B cells (blue), and YFP particles (green) in FALCs from mice fed CFD or HFD for 32 weeks. i–j, Type 1 FALCs were rich in CD31 + blood vessels, T cells and B cells but lacked CD11b + and LYVE + cells (that is LECs and macrophages) and had relatively low uptake of YFP-nanoparticles. k, Type 2 FALCs were rich in T cells, B cells and LYVE-1+ cells (macrophages or LEC) but had no observable lymphatic vessels and moderate uptake of YFP-nanoparticles. l-m, Type 3 FALCs contained, and were in close proximity to, LYVE-1+ lymphatic vessels and CD31 + blood vessels. The lymphatic vessels appeared disorganised and lacked a clear orientation toward a collecting lymphatic vessel. Type 3 FALCs also contained high numbers of T cells, B cells and myeloid cells. The observed overlap in staining between the YFP nanoparticles and CD11b + myeloid cells is most likely due to phagocytosis of the particles by the cells. Scale bars, 100 µm (i-m). (i-m) Representative of images from 2-3 FALCs for n = 5 mice. P values and details of the statistical testing are provided as source data.
