A to E, Rat VSMCs were treated for 24 hours with IL-1β, U46619, or both. TPr antagonist SQ29548 was added 1 hour prior to IL-1β and U46619. A to C, Real-time RT-PCR analysis of mRNA levels of Nox1, VCAM-1, and iNOS. D and E, Western blot analysis of VCAM-1 and iNOS protein expression. Total ERK levels are shown as a reference for equal loading. F and G, RT-PCR analysis of Nox1 and Nox4 mRNA expression in rat VSMCs treated with IL-1β or IL-1β plus U46619 for 16 h (F) or the time as indicated (G). In G, JNK inhibitor SP600125 (1 µmol/L) was added 1 h prior to IL-1β and U46619. H, Real-time RT-PCR analysis of Nox1 mRNA levels in rat VSMCs infected with adenovirus expressing Lac Z or JNK1-dn, and then treated for 24 hours with IL-1β, U46619, or both, as indicated. Bar graphs A to C represent data from 3 separate experiments, and H from 2 separate experiments. The mRNA results were normalized to GAPDH mRNA levels and expressed as fold-changes compared to IL-1β- treated cells.