Extended Data Fig. 3: Lymphatic insufficiency in Apoe^−/− mice reproduces most features of LE pathology.

(a-i) Apolipoprotein E deficient (Apoe^−/−) and WT female mice were switched to a high fat diet at 6 weeks of age. (a) Transport of fluorescent dextran from the rear footpad (site of injection) to the draining lymph node was assessed 10 min after dye injection. Data are expressed as relative fluorescence tracer to WT. n = 11; 13; 15; 20 mice (WT) and 15; 17; 14; 14; mice (Apoe^−/−) in 6–8; 10; 17–26; >30 wks of age. (b) Representative images of mouse footpad across age. (c) Measurement of front footpad thickness. n = 7; 17; 17 mice (WT) and 13; 20; 15 mice (Apoe^−/−) in 10; 17–26; >30 wks of age. (d) Percentage of water content in skin tissues. n = 5; 10; 8 mice (WT) and 8; 9; 7 mice (Apoe^−/−) in 10; 17–26; >30 wks of age. (e) Representative low magnification images of mouse skin stained for CD68 (macrophage), perilipin (adipocyte) and DAPI. Inserts show high magnification. (f) Apoe^−/− mouse skin sections stained for CD3 (T cells), perilipin (adipocyte) and DAPI. (g-i) Gene expression of Tnfα (g), Il6 (h) and Il1β (i). (g) n = 5; 11; 8 mice (WT) in and n = 8; 11; 7 (Apoe^−/−) in 6–8; 10; 17–26; >30 wks of age. (h-i) n = 5; 10; 8 mice (WT) in and n = 8; 12; 7 (Apoe^−/−) in 6–8; 10; 17–26; >30 wks of age. Bar represents mean ± SEM. (a-i) Data are collected from 2–3 independent experiments. P values were calculated using two-way ANOVA with Sidak multiple comparisons. Scale bar = 5 mm in (b), 500 µm and 50 µm for high mag image in (e) and 100 µm in (f). Ep – Epidermis, D – dermis, dAT – dermal AT, PC – Panniculous Carnosus, Fascia - F.