Cell culture grade dimethyl sulfoxide dmso

PTX and DOX (purity >99.5%), and cell culture grade dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO). Drug stock solutions were prepared in DMSO and diluted to desired concentrations such that the DMSO concentration was below 0.5%.

Splenocytes were isolated from either normal or AML mice and treated with rmIL-4 (10 ng/mL, 24hr) or CyPGs (Δ¹²-PGJ₂, 15d-PGJ₂, and Δ¹²-PGJ₃ at 100 nM in sterile PBS for 24hr) and incubated at 37 °C and 5% CO₂. Δ¹²-PGJ₂ and 15d-PGJ₂ were purchased from Cayman Chemical, Ann Arbor, MI, USA. Δ¹²-PGJ₃ was prepared in our laboratory as described previously.^{20 (link)} All three compounds were prepared fresh before use in sterile PBS and concentrations were calculated by liquid chromatography-mass spectrometry (not shown). For mRNA expression analysis, primary AML splenocytes were treated with rmIL-4 (10 ng/mL, 24hr); for protein expression analysis, primary AML splenocytes were treated with HQL79 (50 μM) and GW9662 (10 μM) in the presence of rmIL-4 (50 ng/mL) and incubated at 37°C and 5% CO₂ for 72 h. For the analyses of apoptosis and viability, primary AML splenocytes were treated with rmIL-4 (0, 50, 100 ng/mL) for 24, 48, and 72 h. At the time point of 48 h, GW9662 (10 μM) was added to the treatment of 50 ng/mL rmIL-4. HQL79 and GW9662 were purchased from Cayman Chemical, Ann Arbor, MI, USA, and reconstituted in cell-culture grade dimethyl sulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA).