In humans, thromboxane (TX) A2 signals through two receptor isoforms, thromboxane receptor (TP)alpha and TPbeta, which are transcriptionally regulated by distinct promoters, Prm1 and Prm3, respectively, within the single TP gene. The aim of the current study was to investigate the ability of the endogenous peroxisome proliferator-activated receptor (PPAR)gamma ligand 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2) to regulate expression of the human TP gene and to ascertain its potential effects on the individual TPalpha and TPbeta isoforms. 15d-PGJ2 suppressed Prm3 transcriptional activity and TPbeta mRNA expression in the platelet progenitor megakaryocytic human erythroleukemia (HEL) 92.1.7 cell line but had no effect on Prm1 or Prm2 activity or on TPalpha mRNA expression. 15d-PGJ2 also resulted in reductions in the overall level of TP protein expression and TP-mediated intracellular calcium mobilization in HEL cells. 15d-PGJ2 suppression of Prm3 transcriptional activity and TPbeta mRNA expression was found to occur through a novel mechanism involving direct binding of PPARgamma-retinoic acid X receptor (RXR) heterodimers to a PPARgamma response element (PPRE) composed of two imperfect hexameric direct repeat (DR) sequences centred at -159 and -148, respectively, spaced by five nucleotides (DR5). These data provide direct evidence for the role of PPARgamma in the regulation of human TP gene expression within the vasculature and point to further critical differences in the modes of transcriptional regulation of TPalpha and TPbeta in humans. Moreover, these data highlight a further link between enhanced risk of cardiovascular disease in diabetes mellitus associated with increased synthesis and action of thromboxane A2 (TXA2).