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. 2014 Oct 27;124(12):5352–5367. doi: 10.1172/JCI76561

Figure 6. Neutralizing miR-23a in CTLs mitigates TGF-β–induced immunosuppression.

Figure 6

(A) Purified naive pMel-1 CTLs were activated with anti-CD3/anti-CD28 and the indicated concentrations of TGF-β for 48 hours in vitro, in the presence or absence of 50 nM FAM-tagged anti–miR-23a LNA. The percentage of granzyme B–expressing TCRβ+CD8+FAM– and TCRβ+CD8+FAM+ cells was assessed by flow cytometry. Data shown represent mean ± SEM; n = 8. ***P < 0.001 by 2-way ANOVA and Bonferroni post-test. (B) Mock- and miR-23a decoy–transduced pMel-1 CTLs were cultured with IL-2 for the first 48 hours, then washed and treated with the indicated concentrations of TGF-β for the next 48 hours. CTLs were restimulated with anti-CD3/anti-CD28 (1 μg/ml each) for the final 24 hours, and the percentage of IFN-γ–producing CD8+iRFP+GFP+ cells was assessed by flow cytometry. Data shown represent mean ± SEM; n = 4. *P < 0.05 and **P < 0.01 by 2-way ANOVA and Bonferroni post-test.