FIG. 3.

U21 associates with and relocalizes HLA-G. Control JEG-3 cells (a) and U21-expressing JEG-3 cells (b) were labeled with W6/32, an antibody directed against properly folded class I MHC molecules, followed by an Alexa 488-conjugated secondary antibody. Scale bars = 10 μm. (c) Lysates and U21 immunoprecipitations of control and U21-expressing JEG-3 cells treated with lysosomal protease inhibitors folimycin and leupeptin were immunoblotted with HCA2 to detect HLA-G. The fold increase in signal intensity between bands is indicated by arrows. (d) Crude membrane preparations of U373, JEG-3, and U373 and JEG-3 cells expressing U21 were immunoblotted with MCW50 antibody to U21, as indicated. Three micrograms of U373 crude membranes and 20 μg of JEG-3 crude membranes were immunoblotted, as indicated. The lower band marked by the asterisk is a cross-reacting background band.