AICAR induces apoptosis independently of AMPK. (A) CLL cells were untreated (CT) or treated with 0.5mM AICAR or with 0.5mM phenformin (PHEN) for 24 hours. Viability was measured by flow cytometry, and it is expressed as the percentage of the viability (n = 11). ***P < .005, AICAR-treated versus untreated cells or versus PHEN-treated cells. **P < .01, PHEN-treated versus untreated cells. (B) CLL cells were incubated without (CT) or with 0.5mM AICAR or 100μM A-769662 (A) for 6 and 12 hours. Phospho-ACC and phospho-AMPK were analyzed by Western blot. ERK was used as a control of protein loading. A representative patient is shown (n = 5). (C) CLL cells were untreated (CT) or treated with 0.5mM AICAR or with 100μM A-769662 (A) for 24 hours. Viability was measured by flow cytometry, and it is expressed as the percentage of the viability (n = 18). ***P < .005, AICAR-treated versus untreated or versus A-treated cells. In the boxplots (A,C), the top line represents the 75% quartile; bottom line, the 25% quartile; and middle line, median. The ends of the whiskers represent the minimum and maximum of all the data. (D) WT and Ampkα1^−/− B lymphocytes were incubated for 12 hours with AICAR in a range of concentrations (0.05, 0.1, 0.25, and 0.5mM). Viability was measured by flow cytometry, and it is expressed as the percentage of the viability. Data are mean ± SEM from 3 independent experiments for each genotype. **P < .01, *P < .05, Ampkα1^−/− versus WT B lymphocytes.