| |
Total Splenocytes
|
# of CD4+ DO11.10+
lymphocytes
|
% CD44+
|
|---|
|
STAT6xRAG2 + primed CD4 T cells
|
157 × 106 cells
|
328 cells
|
99.3%
|
|
STAT6xRAG2 + Naïve CD4 T cells
|
350 × 106 cells
|
629 cells
|
99.5%
|
- T cell activation studies were conducted using the protocol mentioned in Figure 2 and materials and methods. Briefly, naïve or in vivo primed CD4+ T cells were adoptively transferred into STAT6xRAG2-/- mice on day 0, followed by OVA/alum immunization of days 1 and 8. Four days later, splenocytes were harvested, single cell suspensions were prepared and cells were counted (column 1). Splenocytes were stimulated with PMA and Ionomycin for 6 h, followed by FACS staining and analysis. Lymphocytes were gated based on forward and side scatter parameters. The CD4+ DO11.10+ population in each transfer group was gated based on double expression of CD4 and KJ126 by each cell (column 2). CD44 expression in these gated cells was examined (column 3). The numbers/percentages in columns 2 and 3 were determined by FACS Analysis. 20,000 events (splenocytes) were collected for each tube/analyte.
