Down-regulation of P2Y₁₁R mRNA expression in matured MoDCs correlates with reduced ATP-mediated arrest of migration. (A) Effect of 100 μM ATP on the migration of MoDCs matured with TNF-α, IFN-α, and PGE₂ for 24 or 48 hours. Migration to CCL21 in the absence of ATP was normalized to 100% (mean ± SEM; n = 7). ^*P < .01. (B) P2Y₁₁R mRNA expression as measured by RT-PCR in immature MoDCs (GM-CSF + IL-4) or those matured overnight with TNF-α + IFN-α + PGE₂, CD40L, or E coli. (C) Quantitative analysis of P2Y₁₁R mRNA expression as measured by quantitative real-time PCR (RT-qPCR). Threshold was exceeded after 24 PCR cycles in immature MoDCs. Immature MoDCs were used as the calibrator to calculate relative P2Y₁₁R mRNA levels in MoDCs matured in the presence of the indicated stimuli. Expression of 18S rRNA within each sample was used for normalization (mean ± SEM; n = 4).