FIGURE 2.
Notch1 AS mice express reduced levels of perforin and granzyme B in CD8+ T cells. A, Splenocytes from Notch1 AS mice were stimulated with anti-CD3ε plus anti-CD28 for 2 days, CD8+ T cells were isolated and we used immunoblotting to analyze whole cell lysates for N1ICD expression (upper bands). Graph (lower) represents band intensity normalized against GAPDH. B, RT-PCR was used to assess perforin and granzyme B mRNA transcript expression in CD8+ T cells from Notch1 AS mice (upper bands). Quantifications of the relative expression of perforin and granzyme B transcripts are represented (lower panels). C, Splenocytes from Notch1 AS mice were isolated and stimulated with anti-CD3ε plus anti-CD28 for 2 days, and intracellular staining was conducted to determine the percentage of CD8+ T cells expressing N1ICD, perforin, and granzyme B; blank peaks indicate isotype control. Data shown represent one of at least three independent replicates.