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Supplementary Figure S1: A compact and simple method of achieving differential transgene expression by exploiting translational readthrough

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posted on 02.03.2022, 13:47 by James Edward Sillibourne, Giulia AgliardiGiulia Agliardi, Matteo Righi, Katerina Smetanova, Grant Rowley, Simon Speller, Abigail Dolor, Katarina Lamb, Christopher Allen, Rajeev Karattil, Farhaan Parekh, Frederick Arce Vargas, Simon Thomas, Shaun-Paul Cordoba, Martin Pule
Supplementary figure S1. TRMs facilitate ultra-low expression of the highly potent cytokine IL-12.A and B) Secretion of IFNγ from splenocytes stimulated with conditioned medium from transduced splenocytes expressing low levels of IL-12 regulated by a translational readthrough motif. A) The data show that IL-12 secretion was undetectable in the culture medium of the transduced splenocytes (levels below the sensitivity of the ELISA). B) When conditioned medium from the transduced splenocytes was transferred to naive splenocytes, IFNγ secretion could be detected by ELISA, suggesting that IL-12 was present in the medium and biologically active. C) Human peripheral blood mononuclear cells were transduced with retroviral constructs encoding human flexi-IL-12, supernatants collected after 72 hours, and cytokine quantification carried out by ultra-sensitive IL-12 ELISA.