Rapamycin does not enhance doxorubicin-induced apoptosis in RelA-hyperexpressing transfectants. (A) Western blotting analysis of p65 (RelA) expression levels in cell lysates obtained from wild-type Jurkat cells and void vector- or RelA-stable transfectants. (B) EMSA analysis of nuclear extracts obtained from Jurkat wild-type cells cultured for 5 hours with and without 5 μM doxorubicin and with or without rapamycin (100 ng/mL), and RelA- or void vector-stable transfectants. (C) Analysis of apoptosis of RelA- or void vector-stable transfectants cultured with and without 5 μM doxorubicin and with and without rapamycin (100 ng/mL). After 24 hours of incubation, cells were harvested and cell death was analyzed by propidium iodide incorporation in flow cytometry. Results are from 4 different experiments, each of which was in triplicate. Error bars indicate standard deviations.