PGE2-induced MoDC migration to CCL19 and CCL21 is PKA and Rho kinase dependent. (A) Titration of PGE2; immature MoDCs were stimulated with sCD40L and titrated concentrations of PGE2. Subsequently, MoDCs were tested for CCL19- and CCL21-triggered migration in transwell assays (left axis) and were analyzed by flow cytometry for the expression of CCR7 (right axis). (B) MoDCs were matured with sCD40L and PGE2 in the presence or absence of the PKA inhibitor H-89 for 48 hours. Subsequently, MoDCs were analyzed by flow cytometry for the expression of CD83 and CCR7. The portion of gated positive cells is given as a percentage, with the mean fluorescence intensity given in parentheses. (C) MoDCs were matured with sCD40L and PGE2 in the presence (▪) or absence (▨) of the PKA inhibitor H-89, or MoDCs matured with sCD40L and PGE2 were preincubated with H-89 for 30 minutes (▦) before they were allowed to migrate in response to CCL21. (D) MoDCs matured with sCD40L and PGE2 were left untreated (▨) or were pretreated with the Rho kinase inhibitor Y-27632 (▦) and were analyzed in a migration assay in response to CCL21. Results from 1 of 3 representative experiments are shown. Error bars indicate SEM and asterisks indicate a significant reduction.