FIG.4.

RAG-1 and RAG-2 cooperate in localizing the V(D)J recombinase. NIH 3T3 cells were transfected with CFP-RAG-2 constructs (green pseudo-color) and YFP-RAG-1 constructs (red pseudo-color) and assayed by fluorescence microscopy. Representative cells are shown. (A) Wild-type YFP-RAG-1 was transfected either alone or in combination with CFP-fused RAG-2 proteins. DAPI, RAG-1 (YFP), or merged DAPI and YFP signals are displayed for transfection with RAG-1 alone (top panel). Coexpression patterns of YFP-RAG-1 and CFP fusions containing full-length RAG-2(1-527) or RAG-2(499/508A₁₀) are displayed in the lower panel. In each case, RAG-2, RAG-1, and merged signals are shown. Numbers indicate the percentages of cells positive for RAG-1 and RAG-2 and demonstrating RAG-2 localization patterns similar to those shown. The graph at right displays the localization pattern of RAG-2(499/508A₁₀) in the presence (solid bars) or absence (hatched bars) of wild-type RAG-1. (B) Transfection of YFP-RAG-1(BV) alone (top panel) or with CFP-RAG-2 (lower panel), displayed as in panel A. Numbers indicate the percentages of cells positive for RAG-1 and RAG-2 and demonstrating RAG-2 localization patterns similar to those shown. The graph at right displays localization pattern of RAG-2(499/508A₁₀) in the presence (solid bars) or absence (hatched bars) of RAG-1(BV). (C) Transfection of YFP-RAG-1(1/2BIV) either alone (top panel) or with CFP fused to wild-type RAG-2 (lower panel), displayed as in panel A. Numbers indicate the percentages of cells positive for RAG-1 and RAG-2 and demonstrating RAG-1 localization patterns similar to those shown. The graph at right displays localization pattern of RAG-1(1/2 BIV) in the presence (solid bars) or absence (hatched bars) of wild-type RAG-2.