FIGURE 3.

BMDM-mediated trans-infection of MLV-GFP efficiently targets activated primary B-cells. Splenocytes from WT mice were activated with either LPS (A) or ConA/IL-2 (B) for 3 days and then seeded either alone or onto BMDM from WT or KO mice in a ratio of 1:1. Cultures were challenged with MLV-GFP (m.o.i. 0.2) and splenocytes analyzed for GFP expression 48 h later by flow cytometry. A and B, effect of activation protocols, and the identity of MLV-GFP-infected splenocytes was determined by co-staining for the lineage markers CD3 (T-cells) and CD19 (B-cells). R1 identifies the viable cells. R2* (red box) identifies the MLV-GFP-positive cells and R2 (black box) all viable cells. Dots plots in the lower panels depict the respective mCD3/mCD19 stainings. C, chart bars depict the arithmetic means ± S.D. of the percentage of GFP-positive, viable cells from analyses performed in triplicate. Experiments were performed using 2–3 KO or WT mice and were repeated at least twice showing similar results.