Platelet production from Meg-01 cells by 15d-PGJ2 is independent of PPARγ. (A) Cells were transiently transfected with a PPRE-luciferase construct and treated with either 10 μM 15d-PGJ2, 9,10 dihydro-15d-PGJ2, or rosiglitazone. Twenty-four hours after ligand treatment, a luciferase assay was performed. Cells treated with PPARγ ligands had increased luciferase activity compared with the untreated cells. (B) Meg-01 cells were treated with DMSO (vehicle control) or with 10 μM rosiglitazone, 15d-PGD2, 9,10 dihydro-15d-PGJ2, 15d-PGJ2, or PGJ2 and platelet number was assessed by flow cytometry. Results are presented as mean plus or minus SD (P < .01). (C) Western blot showing that cells infected with PPARγ-siRNA have 66% less PPARγ protein compared with cells infected with the control (con) virus. (D) Cells either infected with lentivirus PPARγ-siRNA or pretreated for 2 hours with 100 nM GW9662, an irreversible PPARγ antagonist, were treated with 15d-PGJ2 for 24 hours. Platelet production was assessed by flow cytometry. Results are presented as mean plus or minus SD (P < .01, n = 3).
