Figure 4.
FcRn-mediated recycling in a cellular system. A) A schematic illustration of the HERA protocol. Briefly, HMEC-1 cells overexpressing human FcRn are seeded one day prior to the experiment. Cells are then washed and starved before samples are added and incubated for 4 hours. Before incubation ON, the medium is replaced. Recycling was defined as amount of protein taken up after 4 hours of incubation and further released back into the medium after ON incubation . The amounts of proteins present in the medium are then quantified in ELISA. The figure was made with BioRender. B) IgA1, IgA1-LCABD and IgA1-HCABD pre-incubated with human albumin were added to the cells. The amounts of the human IgA1 variants in collected medium samples were quantified by ELISA. The numbers given represent duplicates in ELISA of triplicates in HERA (SD). ns > 0.05, * = p 0.0136, **** = p < .0001, by two-tailed analysis using unpaired T-test (df = 10, t = 2.99, 11.05)