Extended Data Fig. 7: Amelioration of lymphedema pathological features in Apoe^−/− mice by HDL and ApoA-I treatment.

(a) Analysis of commercially available vLDL, LDL, HDL and albumin by fast performance liquid chromatography (FPLC) indicating that HDL was eluted at column volume 1.8 ml. (b) Similar to commercial HDL, FPLC elution profile of HDL isolated from healthy human plasma was eluted at column volume 1.8 ml. (c) Analysis of HDL isolated from healthy human plasma by SDS-PAGE (12%) and proteins were detected by Coomassie blue staining. Commercial HDL and albumin were used as positive controls. Isolated HDL from various donors indicated negligible albumin contamination, confirming the efficiency of our HDL isolation method. (d) Immunoblotting of apoA-I from different healthy donors’ HDL. Commercial HDL and albumin were used as positive and negative controls respectively. (a-d) The same findings were observed in 2 repeated experiments. (e) Experimental setup for (f-u). Created in BioRender. Lim, H. (2026) https://BioRender.com/pzqny3l. Apoe^−/− mice (6–8 weeks old) were switched to high fat diet. At around 17 weeks old, mice received intravenous injection of HDL, ApoA-I and HOCI:ApoA-I (20 mg/kg) or PBS (vehicle) for 7 consecutive days. (f) Total plasma cholesterol and (g) HDL cholesterol in mg/dl. n = 7 mice (Veh) and 7 mice (HDL). (h) Lipid stain in mouse skin using Oil Red O in vehicle and HDL treated Apoe^−/− mice. (i) Total cholesterol in mouse skin with n = 4 mice per group. (j) Skin sections from vehicle, ApoAI and HOCI:ApoAI treated Apoe^−/− mice stained with Oil Red O. (k) Mouse skin sections stained for free cholesterol using filipin. (l-m) Free cholesterol content was quantified in the dermis (l) and dermal AT (m) with n = 6 mice (Veh), 5 mice (ApoAI) and 5 mice (HOCI:ApoAI). (n) Measurement of front footpad thickness with n = 4 mice (Veh), 6 mice (ApoAI) and 5 mice (HOCI:ApoAI). (p) Representative images of mouse skin stained with haematoxylin and eosin. (o, q-r) Adipocyte density (o), mean adipocyte area (q) and frequency of small (≤2000 µm²), medium (2001–4000 µm²), large (≥4001 µm²) adipocyte size distribution (r) with n = 6 mice (Veh), 5 mice (ApoAI) and 5 mice (HOCI:ApoAI). (s) Mouse skin stained for collagen using picrosirius red. (t-u) Dermal (t) and dermal AT collagen (u) area were quantified. n = 6 mice (Veh), 6 mice (ApoAI) and 5 mice (HOCI:ApoAI). (f-u) Data are pooled from 2 independent experiments. Bar represents mean ± SEM. P values were calculated using two-tailed Mann–Whitney U-tests in (f, g and i) and one-way non-parametric ANOVA using Dunn’s multiple comparison test in (l-o, q-r and t-u). Scale bar = 200 µm. Ep – Epidermis, D – dermis, dAT – dermal AT, M - muscle.