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. Author manuscript; available in PMC: 2011 Mar 1.
Published in final edited form as: J Immunol. 2010 Feb 5;184(5):2401–2409. doi: 10.4049/jimmunol.0902448

FIGURE 7.

FIGURE 7

Bcl-2 family member expression in agonist and variant epitope-stimulated CD8+ CL-4 T cells. CD8+ CL-4 T cells were stimulated in vitro with 10−6 M PR/8 agonist or Japan variant viral epitopes. A, Cells were analyzed for intracellular Bcl-2 and Bcl-xL expression by flow cytometry. Histograms represent live-gated Thy1.2+ cells. Shaded histograms represent isotype controls. Numbers indicate mean fluorescence intensity. B, At day 1 and day 2 poststimulation, Thy1.2+ cells were purified from culture and lysed in lysis buffer. Equivalent numbers of cells were loaded onto 12% SDS-PAGE gels, and separated proteins were transferred to polyvinylidene difluoride. Proteins were visualized by staining blots with Abs to Bim or β-actin, followed by staining with anti-rabbit IgG-HRP Abs, and developed with ECL substrate. Intensities of Bim-specific bands were measured by densitometry and expressed as arbitrary units normalized to the actin loading controls.