Effects of RAPTOR and RICTOR knockdown. (A-B) Seventy-two hours after transfection with shRNAs targeting RAPTOR (shRap) or RICTOR (shRic) along with plasmid encoding EGFP-histone H2B (to mark transfected cells), Jurkat cells were harvested for immunoblotting or stained with allophycocyanin-annexin V and analyzed by flow cytometry for annexin V binding to EGFP-histone H2B⁺ cells. (C) Cells were transfected with shRNAs, incubated for 12 hours, and treated with diluent or OSI-027 for 72 hours before annexin V staining. Error bars in panels B, C, and all subsequent figures indicate mean ± standard error of the mean of 3 independent assays unless otherwise stated. In panels B and C, *P < .05 relative to diluent-treated cells. In panel C, **P < .05 compared with cells treated with control shRNA and 20 µM OSI-027. All P values in this and subsequent figures are corrected for multiple comparisons.