fig4

Figure 4. HMEV protein evidence after casein micelle removal treatment. Proteomic analysis of HMEVs isolated after casein micelle disaggregation/removal by sodium citrate, EDTA, acetic acid, and chymosin treatments [full results in Supplementary Table 1]. (A) Fold-changes of 87 HMEV proteins as compared to the untreated condition (details in Supplementary Table 2). Color codes: blue, common and specific HMEV markers; green, highly abundant soluble proteins in human milk; black, caseins; (B) The average protein intensities of selected HMEV markers, total caseins (alpha-, beta-, kappa-caseins), and the ratio of caseins per CD9; (C) Western immunoblot demonstrated that different casein micelle removal methods exhibited different capabilities of casein depletion and HMEV enrichment (full-length blots of the cropped images were provided in Supplementary Figure 2). BTN1A1: butyrophilin 1A1; BSSL: bile salt-stimulated lipase; LALBA: alpha-lactalbumin; LF: lactoferrin; MFGE8: milk fat globule-epidermal growth factor 8 (also known as lactadherin); MUC1: mucin 1; NaCit: sodium citrate; TNC: tenascin; XDH: xanthine dehydrogenase. EDTA: ethylenediaminetetraacetic acid.