CC and CXC chemokine production in Fas-ligated DCs. (A) CC (MCP-1, MIP-1α, MIP-1β, RANTES, TARC) and CXC (MIP-2) chemokine production by DCs from C57BL/6J (DC) or lpr (lprDC) mice was measured by ELISA following stimulation of DCs with 0.5 μg/mL Jo-2 (Jo-2), isotype antibody (Iso), 5 μg/mL rmFasL in the presence of 10 μg/mL anti-6 × histidine antibody (FH), or 0.5 μg/mL LPS (LPS) for 18 hours. Unstimulated DCs were used as a control (Ctrl). (B) The kinetics of MIP-1α and MIP-2 production by DCs following stimulation with 0.5 μg/mL Jo-2. (C) MIP-1α and MIP-2 production induced by various concentrations of Jo-2 (0.01-5 μg/mL). (D) Chemokine mRNA expression in DCs stimulated with medium alone (Ctrl), Jo-2, isotype antibody (Iso), or LPS at indicated time points. Results are representative of 3 independent experiments. (E) DCs were incubated with 0.5 μg/mL cycloheximide (CHX) for 30 minutes, and then stimulated with Jo-2, isotype antibody, or LPS for 18 hours, and MIP-1α and MIP-2 levels in supernatant were measured by ELISA. (F) The concentrations of MIP-1α and MIP-2 in the lysates of resting and stimulated DCs at different times were detected. (G) DCs were incubated with CHX for 30 minutes prior to stimulation with Jo-2 for 12 hours. Then we washed this DC culture and restimulated with Jo-2 before testing for the concentrations of MIP-1α and MIP-2 in the supernatant. Untreated and CHX-treated DCs were used as a control. ^*P < .05. Values in panels A-C and E-F are expressed as means ± standard deviation.