Figure 2.

TMEP expression, DC maturation and cell viability analyses after RNA or DNA transfection of MDDCs from HD by flow cytometry. MDDCs were mock-transfected (w/o) or transfected with 10 µg (electroporation) or 15 µg (lipofection) of RNA-TMEP, RNA-TMEPmod or DNA-TMEP. MDDCs lipofected with 15 µg of DNA-ø were used as a control. At 6, 24 and 48 h post-transfection, cells were collected and processed for flow cytometry analysis as described in Materials and Methods. (A,B) FLAG expression was analyzed using 5 µg/mL of the monoclonal antibody anti-FLAG M2. (C,D) DC maturation of MDDCs electroporated (C) or lipofected (D) was analyzed using specific antibodies for CD86, CD80 and CD83 markers. (E) Cell viability of MDDCs was determined using Annexin V-PE/7-AAD.