Figure 1.

Functional expression of FcRY in IMCD cells. (A) Anti-FcRY Western blot of detergent lysates from FcRY-transfected IMCD and untransfected IMCD cells using a polyclonal ascites raised against the FcRY ectodomain. (B) Confocal immunofluorescence of FcRY-IMCD and untransfected IMCD cells stained with anti-FcRY polyclonal ascites. Images were collected with the same laser power, exposure times, and detector settings. (C) Confocal images of FcRY-IMCD cells incubated with 500 nM Alexa 488–labeled IgY or FcY at pH 6 and 8. Internalized ligand was seen only when incubated at acidic pH. (D) Confocal images demonstrating colocalization of FcRY and IgY. Filter-grown monolayers of FcRY-IMCD cells were incubated with Alexa 488–labeled IgY at pH 6 and stained with anti-FcRY. Optical sections were taken ∼1 μm below the tight junctions (top panels) or ∼6 μm below the tight junctions (bottom panels). Scale bars, 10 μm.