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Figure 4 | Biology Direct

Figure 4

From: Critical role for BIM in T cell receptor restimulation-induced death

Figure 4

Fas and Bim cooperate in driving TCR-induced apoptosis of murine T cells. (A) Activated splenic T cells from wild-type (WT), lpr, or bim-/- mice were restimulated with platebound anti-CD3 for 24 h. Percent cell loss was calculated in triplicate by PI exclusion. (B) Lysates from splenic T cells from the indicated genetic backgrounds left untreated or restimulated with platebound anti-CD3 for 6 h were immunoblotted for Bim isoform expression. β-actin serves as a loading control; asterisk indicates non-specific band. (C, D) CD4+ and CD8+ T cells were purified from activated WT, lpr, or bim-/- splenocytes and restimulated with platebound anti-CD3 for 24 h. Percent cell loss was calculated in triplicate by PI exclusion. (E) Splenic T cells from WT or lpr mice were stimulated for 48 h with platebound anti-CD3/anti-CD28, washed, and transfected with NS or Bim-specific siRNA. Three days post-transfection, cells were restimulated with 100 ng/ml platebound anti-CD3; percent cell loss was calculated in triplicate by PI exclusion. Differences in apoptosis sensitivity (relative to NS-treated WT cells) were statistically significant (p < 0.04). Lysates made from cells three days post-transfection were assessed for Bim knockdown by immunoblotting, right.

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