AML blasts induce adipocyte lipolysis. (A) Primary AML blasts or nonmalignant CD34⁺ cells were cultured alone or in coculture with adipocytes for 24 hours. Media were removed and used to detect free FA and glycerol. Data are represented as mean ± standard deviation. (B) AML blasts incubated with media supplemented with BSA or media supplemented with 100 µM of oleate-BSA conjugate for 2 days and AML blasts counted using flow cytometry and Trypan blue exclusion (n = 4). The line through the data indicates the median. (C) Immunoblot for pHSL from adipocytes cultured with and without AML blasts (n = 4). Blots were reprobed for total HSL and β-actin to show equal sample loading. (D) Primary AML blasts in monoculture or cultured with adipocytes (adip) or BMSC with and without treatment with acipomox (APX) (10 μM) for 72 hours. AML blasts were counted using flow cytometry and Trypan blue exclusion (n = 4). The Mann-Whitney U test was used to determine statistical significance between treatment groups. Data are represented as mean ± standard deviation. (E-F) AML blasts cultured on adipocytes (ADI; with and without acipomox, 10 μM) or BMSC that had been preincubated with fluorescent DDA for 24 hours (n = 4). Blasts were analyzed for uptake of the fluorescent DDA by flow cytometry. Data are represented as mean ± standard deviation. (E) The red line is AML cultured on BMSC; the black line is AML cultured on adipocytes, and the blue line is AML cultured on adipocytes treated with acipomox. *P < .05. FFA, free fatty acid.