Fig. 4: Fc-MST-HN shows higher levels of intracellular FcRn occupancy compared with IgG1-MST-HN.
From: The Fab region of IgG impairs the internalization pathway of FcRn upon Fc engagement
a Histograms of a representative experiment comparing the binding of AF650-labeled anti-FcRn Fab to FcRn in HEK-FcRn-GFP cells after pre-loading cells with indicated IgG variants at 37 °C for 2 h and subsequently removing surface-bound IgG by extensive washing. b Anti-FcRn Fab-AF650 staining of cells plotted as percent gMFI in the presence of indicated inhibitors compared with untreated cells. c Intracellular accumulation of equally AF405-labeled molecules in HEK-FcRn-GFP cells after pre-loading and washing the cells as in (a) as measured using flow cytometry. Data represent histograms from a representative experiment, with indicated numbers showing gMFIs averaged from measured duplicates from the same experiment in (a), with aggregated data presented as mean values from three independent experiments in (b) and (c). Error bars indicate standard deviations. Statistical analysis was performed using a two-way ANOVA (Sidak’s multiple-comparisons test), and statistically significant differences are indicated by asterisks **<0.01, ***<0.001, ****<0.0001. ns not significant.
