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For immunofluorescence analysis, the sections were heated in a microwave oven in citrate buffer, after cooling, blocked with 5% BSA in PBS and 0.5% (wt/vol) Triton X-100 for 30 min at room temperature, and subsequently incubated with primary antibodies at 4 C overnight and secondary antibodies for 1 h at room temperature, followed by washing and staining with DAPI (Solarbio, S2110)

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The human albumin variants were expressed in a serum-free transient Expi293 expression system, and all were secreted at comparable quantities to that of WT albumin (2546 mg/L) (Supplementary Fig

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