Figure 2. Effects of FcRn antagonists on recycling of HSA by HEK293-hFcRn-GFP cells.

HEK293-hFcRn-GFP cells were incubated with 50 nM FcRn antagonist or medium for 24 hours. The cells were then incubated in serum-free medium for 2 hours, pulsed with 250 μg/mL AF647-labeled HSA (HSA-AF647) in serum-free medium for 1 hour, washed, and chased in serum-free medium for 0 (C-) or 30 minutes (C+) at 37°C in a 5% CO₂ incubator. The cell-associated HSA-AF647 levels following the indicated treatments were determined using flow cytometry. (A) Schematic illustration of HSA recycling assay. (B) Data normalized against pulse-only levels (represented as 100%). These data are combined from 2 independent experiments, with triplicate samples in each experiment. One-way ANOVA was used for statistical analysis. Statistically significant differences are shown as *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. Error bars indicate the standard deviation of the mean.