Figure 1

Expression of tumor-associated HPV E6 and E7 antigens by bone marrow-derived DCs after RNA electroporation. (A) Gel-like image obtained from 10 independent samples of TC-1 RNA analyzed by the Agilent bioanalyzer. (B) Electropherogram of sample 1 from A showing ribosomal RNA peaks. (C) Flow cytometry analysis of intracellular E7 expression in CD11c⁺ cells 24 hours after electroporation with TC-1 RNA at different voltage and capacitance conditions. Electroporation was performed with 25 μg TC-1 RNA/10⁶ DCs (grey): mock electroporated (white). (D) Flow cytometry analysis of DC viability 24 hours after electroporation at different conditions of voltage and capacitance. Electroporation was performed with 25 μg TC-1 RNA/10⁶ DCs. X-axis reflects propidium iodide (PI) staining. (E) Flow cytometry analysis of intracellular E6 and E7 expression in CD11c⁺ cells 24 hours after electroporation with different amounts of TC-1 RNA at 300V and 150 μF. The experiment was repeated two times with similar results. (F) Flow cytometry analysis of intracellular E6 and E7 expression in CD11c⁺ cells 4 days after electroporation with 50 μg of TC-1 RNA/10⁶ DCs at 300V and 150 μF. RNA electroporated (white); mock electroporated (grey). (G) Immunofluorescence of DCs stained with anti-HPV E7 antibody 24 hours after electroporation with TC-1 cell RNA or mock electroporated. Cells were counterstained with DAPI. 200X magnification. All experiments were repeated twice with similar results.