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Figure 3 Effects of CB₁ Antagonists on DOX-Induced Cell Death and Apoptosis In Vitro

(A) Effects of CB₁ antagonists on cell viability measured by XTT assays. AM281 and rimonabant (1 µmol/l) prevent cell death induced by 1 or 5 µmol/l of DOX. *p < 0.05 versus control group; #p < 0.05 versus DOX (n = 4). (B) Effects of CB₁ antagonists on the early apoptosis marker fluorescent annexin V conjugate (Annexin V-FITC) and cell death detection dye propidium iodide (PI) measured by flow cytometric analysis of H9c2. Representative data from 3 experiments analyzed. (C) Effects of CB₁ antagonists on active caspase expression (green) and nuclear staining pattern by Hoechst 33342 dye (blue). Please also note that some of the nuclei are fragmented in DOX-treated cells (indicative of late apoptosis), but not in CB₁ antagonist-treated or control cells. Representative data from at least 15 experiments analyzed. Abbreviations as in Figure 1.