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. 2015 Jun 24;5(1):e1062967. doi: 10.1080/2162402X.2015.1062967

Figure 6.

Figure 6.

CD137 silencing in CT26 tumor cells gives rise to more immunogenic variants. CT26 were stably transfected with lentiviral vectors to express a shRNA targeting CD137 (shCD137) or a scrambled control (shControl). (A) Transfectants were cultured for 36h in normoxia or 1% O2 and analyzed by flow cytometry to quantitatively determine the expression of surface CD137. (B) Quantitative RT-PCR analysis of total and transmembrane CD137 mRNA in the indicated transfectants under hypoxic or normoxic conditions as in A. (C) BALB/c wild type immunocompetent mice were subcutaneously inoculated with 5×105 cells of indicated transfectants and tumor sizes were individually followed over time. Tumor cells had been pre-exposed in every case to 1% O2 for 36h. The fraction of mice spontaneously rejecting their tumors is given in each graph. When indicated, BALB/c mice were mAb-depleted of CD8+T cells. (D) mean±SEM and Statistical comparisons of experiments in C whose results are representative of two independent experiments. (E) Tumor growth of the indicated transfectants in immunodeficient Rag2IL2Rγ−/− mice. Tm: transmembrane.