Figure 7. LAG-3 expression has a minor impact on cell proliferation and apoptotic pathways.

2×10³ Thy1.1+Ly5a− WT P14 cells and 2×10³ Thy1.1−Ly5a+ LAG-3−/− P14 cells mice were adoptively transferred into the same WT B6 mice (Thy1.1−Ly5a−); 4 days later the mice were infected with LCMV-Armstrong or Clone13. (A) The gMFI of Bcl-2 and Bim on WT and LAG-3−/− P14 cells at days 7 and 13 during Clone13 infection. (B) The Bim:Bcl-2 ratio within each cell type at day 13 of Clone13 infection. (C and D) Splenocytes were isolated on days 7 or 13 and cultured in vitro for 6 hours. The frequency of AnnexinV+ (C) and activated Caspase 3 (D) P14 cells after the in vitro culture. (E) The frequency of Ki-67+ splenic WT and LAG-3−/− P14 cells at various times after infection. (F) Illustration showing the design of the in vivo proliferation assay. At 8 days post-infection, CD8+ T cells were isolated by MACS negative selection, CFSE labeled, and re-transferred to day 8 Clone13 infected mice that had not previously received P14 cells; CFSE dilution was analyzed 3 days later. (G) Examples of CFSE dilution by WT and LAG3−/− P14 cells (left) and the total frequency of cells that had divided (right), with the lines indicating the paired analyses. The data in panels A and B represent 11 mice from 4 independent experiments. The data in panel C, D, and G represent 6 mice from 2 independent experiments. The data in panel E represent 9 mice from 3 independent experiments.