Fig. 2.

C5L2 has a primarily intracellular location in transfected RBL cells. (A) The expression pattern of receptor in RBL cells transfected with C5aR, wild type or mutant C5L2 (NPXXY) was measured by incubating permeabilized and intact cells with anti-receptor antibodies. The percentage surface expression was calculated as the ratio of antibody binding to intact cells to that in permeabilized cells. The data shown are the mean ± S.E.M. of three to four separate determinations performed in duplicate. Significantly different by ANOVA with Bonferroni post-test, *p < 0.05; ns: not significant. (B) Intracellular and surface receptor expression was visualized in RBL cells transfected adhered to coverslips, incubated with antibody specific for the N-terminus of C5aR or the N-terminal HA tag of C5L2 with or without a permeabilization step; nuclei are counter-stained in blue. Bar equal to 10 μm. (C) CHO cells transfected with GFP tagged constructs of C5aR or C5L2 mutants were adhered to coverslips. Bar equal to 10 μm.