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. 2011 Mar 17;18(8):750–764. doi: 10.1038/gt.2011.15

Figure 1.

Figure 1

Vector design and immunophenotypic analyses: (a) tricistronic vectors: diagram representing the tricistronic vectors simultaneously co-expressing suicide gene sr39HSV-TK, GM-CSF and IL-4 with the immediate early CMV promoter or with MHCII promoter. The tricistronic construct contain two 2A-like sequences (the porcine teschovirus (P2A) and the Thosea asigna virus (T2A) upstream of mouse GM-CSF and mouse IL-4 open-reading frames. The amino acid sequences of the two heterologous 2A elements are indicated, and the cleavage sites for the protein products are indicated by the arrow. Monocistronic vectors: diagram representing the monocistronic vectors encoding melanoma-associated tumor antigen TRP2 or fLUC. (b) Schematic representation of SMART-DC production and analyses. (c) May–Grunwald Giemsa stain of cytospin preparations, performed at each time point showing the typical morphology of DCs. Representative data from three independent experiments are shown. (d) Viability at early time points was evaluated as total cell counts (obtained by trypan blue exclusion staining) and as percentage of viable CD11c+ cells (negative for Annexin V and 7-AAD staining).