FIGURE 3.
Characterization of the Heat-Inducible Plasmid. A) Schematic depicting the sTRAIL-EGFP plasmid (i), which expresses a sTRAIL-EGFP fusion that is constitutively activate, and the HSP-sTRAIL plasmid (ii), which expresses the same sTRAIL-EGFP fusion under the control of a heat-inducible HSP70B’ promoter. B) RT-PCR demonstrating the successful synthesis of the sTRAIL-EGFP plasmid, which was transfected into A2780 ovarian cancer cells. C) Proof-of-concept demonstrating that the HSP-sTRAIL plasmid can be specifically activated by heat (1 hour at 41°C in a water bath) as seen via fluorescence imaging due to fusion of TRAIL with EGFP. Scale bar = 50 µm. D) Confirmation of heat-specific TRAIL activation was obtained using qPCR (*p < 0.05) and was normalized to transfected cells that were incubated at 37°C. GAPDH was used as the housekeeping gene.