Journal of Immunology Research / 2016 / Article / Fig 3

Research Article

Transcriptional Targeting of Mature Dendritic Cells with Adenoviral Vectors via a Modular Promoter System for Antigen Expression and Functional Manipulation

Figure 3

The constitutively active mHSF1 induces specific transgene expression in HeLa cells and human DCs. (a, b) HeLa cells were transfected with 0.7 μg DNA of plasmid vectors expressing MelanA/MART-1 (“MelA”), BclxL or/and IL-12 under the control of the Hsp70B′−29/−242 heat shock response element as indicated. The next day, cells were either left untreated (“Mock”) or transduced with adenoviruses Ad5Luc1 or Ad5mHSF1 at 300 TCID50/cell. (c, d) Human immature DCs were cotransduced in the presence of the maturation cytokine cocktail with adenoviruses as indicated at a total amount of 500 TCID50/cell. (a–d) Twenty-four hours later, cell lysates were analyzed for BclxL and MelanA expression by Western Blot (a, c) and cell culture supernatants for content of IL-12p40 and IL-12p70 by ELISA (b, d). (a and c) Showing one representative experiment out of three; (b and d) data are mean ± SEM of three independent experiments with different donors. , , and n.s.: not significant (); bars without annotation are not significant (); one-way ANOVA with Bonferroni’s Multiple Comparison post hoc test.

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