Figure 1: Mesothelin-targeting increases the bioactivity of soluble TR3. | Scientific Reports

Figure 1: Mesothelin-targeting increases the bioactivity of soluble TR3.

From: Membrane-proximal TRAIL species are incapable of inducing short circuit apoptosis signaling: Implications for drug development and basic cytokine biology

Figure 1

(a) Schematic representation of recombinant TR3 forms. The functional domain of secreted wild-type TRAIL has been genetically linked, resulting in the TRAIL-based drug platform TR3 (left). TR3 was subsequently fused at its N-terminus with scFv-SS, resulting in the mesothelin-targeted TR3 variant SS-TR3 (right). (b) Western blot analysis of recombinant TR3 forms produced in HEK293T cells (anti-TRAIL pAb). Supernatant from mock transfected cells was used as a control (lane 1). (c) Schematic representation of FLAG-tagged, mature human mesothelin inserted into the membrane via a glycosylphosphatidyl (GPI) anchor. Western blot analysis of Jurkat cells infected with a retroviral vector encoding FLAG-tagged human mesothelin (~40 kDa). (d) Anti-FLAG immunostaining confirms expression of the biomarker on Jurkat cells using flow cytometry. (e) Jurkat-WT and (f) ~5% Jurkat-Meso cells were treated with increasing concentrations of TR3 and SS-TR3 and cell viability was determined (n = 3, mean ± SEM; ns, not significant, *P < 0.025, t-test).

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