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. 2012 Sep 25;2012:745962. doi: 10.1155/2012/745962

Figure 5.

Figure 5

HSP70 inhibitor blocks both endogenous- and cross-presentation. (a) DC2.4 cells were transfected with a plasmid containing cDNA encoding OVA by electroporation and immediately incubated with the indicated inhibitors. Three hours later, the cells were fixed and incubated with OTI-CD8+ T cells for 24 hours (left panels). Also, DC2.4 cells transfected with an empty plasmid were pulsed with a suboptimal dose of OVA(257–264) peptide (10−9 M) for three hours in the presence of inhibitors (20 μM each), fixed and were used as positive controls in the same assay (right panel). (b) DC2.4 cells were incubated with 0.5 mg/mL OVA in the presence of the indicated inhibitors for three hours and fixed. The cells were washed and incubated with OTI-CD8+ T cells for 24 hours (left panel). Also, DC2.4 cells were pulsed with 0.5 mg/mL OVA in the presence of the indicated inhibitors (20 μM each) for three hours, fixed and were used as positive controls in the same assay (right panel). The production of secreted IFNγ by T cells in the supernatants was evaluated by ELISA.*P < 0.05; **P < 0.01; ***P < 0.001. The results were confirmed in at least two independent experiments.