American Society of Hematology

Fig. 5.

$Fig. 5. Enhanced Bcl-2 expression and cell survival after stem cell gene correction. / (A) Bcl-2 expression levels in thymocyte fractions. Thymocytes were stained for surface CD4 and CD8 expression, then stained intracellularly with either isotypic control mAb (gray histograms) or antimouse Bcl-2 mAb (open histograms). Shown are the results analyzed by gating CD4−/CD8− (DN) cells, CD4+/CD8+ (DP), or either CD4+ or CD8+ cells. Representative results of 2 independent experiments are presented. (B) In vitro assessment of thymocyte cell death. After 24 hours of culture, cells were stained for CD4 and CD8, then incubated with 7-AAD. The percentage of 7-AAD+ cells was determined in each thymocyte fraction by gating on CD4−/CD8−, CD4+/CD8+, CD4+, and CD8+ cells. Shown are the mean values of 2 independent experiments. ▪, γc+, untreated normal mice; ■, XSCID, untreated Δγc+-XSCID mice; ░, mγc-BMT, Δγc+-XSCID mice transplanted with mγc-transduced BM cells.$

Enhanced Bcl-2 expression and cell survival after stem cell gene correction.

(A) Bcl-2 expression levels in thymocyte fractions. Thymocytes were stained for surface CD4 and CD8 expression, then stained intracellularly with either isotypic control mAb (gray histograms) or antimouse Bcl-2 mAb (open histograms). Shown are the results analyzed by gating CD4⁻/CD8⁻ (DN) cells, CD4⁺/CD8⁺ (DP), or either CD4⁺ or CD8⁺ cells. Representative results of 2 independent experiments are presented. (B) In vitro assessment of thymocyte cell death. After 24 hours of culture, cells were stained for CD4 and CD8, then incubated with 7-AAD. The percentage of 7-AAD⁺ cells was determined in each thymocyte fraction by gating on CD4⁻/CD8⁻, CD4⁺/CD8⁺, CD4⁺, and CD8⁺ cells. Shown are the mean values of 2 independent experiments. ▪, γc⁺, untreated normal mice; ■, XSCID, untreated Δγc⁺-XSCID mice; ░, mγc-BMT, Δγc⁺-XSCID mice transplanted with mγc-transduced BM cells.

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