Electroporation of 3d mDC and 7d mDC with long MART-1/Melan-A peptide and MART-1/Melan-A-encoding ivt RNA. (A) 3d mDC and 7d mDC were electroporated (250 V, 150 μF) with 1 μg, 5 μg and 10 μg long MART-1/Melan-A peptide. After 24 h incubation at 37°C and 5% CO2, the DC were cocultured with A42 CTL for 24 h. (B) 2d iDC, 3d mDC and 7d mDC were electroporated (250 V, 150 μF) with 24 μg MART-1/Melan-A ivt RNA, incubated at 37°C for 24 h and cocultured with A42 CTL for 24 h (n = 3). (C) 3d mDC and 7d mDC were electroporated with 12 μg MART-1/Melan-A ivt RNA at different electroporation conditions (250 V, 150 μF and 300 V, 300 μF), respectively. 3 h after electroporation, mDC were stained intracellularly with a MART-1/Melan-A-specific antibody and analyzed by flow cytometry (n = 2). (D) 24 h after electroporation with MART-1/Melan-A ivt RNA, DC were cocultured with A42 CTL for 24 h (n = 2). The IFN-γ release of the A42 CTL was measured by IFN-γ-ELISA. The bars in A, B and D show mean values of triplicates with standard deviations (Rec.: recovery; n.d.: not detected).