Fig. 3: FTO-deficient effector CD8+ T cells have increased cell apoptosis.
From: FTO controls CD8+ T cell survival and effector response by modulating m6A methylation of Fas
A Splenocytes from WT (Ftofl/flCD4-Cre−) and FTO KO (Ftofl/flCD4-Cre+) mice were stimulated with anti-CD3/CD28 antibodies for 24 h in the presence of Z-VAD to measure Annexin V and 7AAD expression in CD8+ T cells (n = 3). B GO analysis of DEGs from RNA-seq data depicting the upregulated and downregulated signaling pathways in FTO KO CD8+ T cells compared to WT control. C Hallmark gene sets associated signaling pathways were enriched in FTO KO CD8+ T cells. D Representative flow cytometry plots and the mean fluorescence intensity (MFI) of Bcl-2 expression in WT and FTO KO donor-derived (OT-1+) CD8+ T cells in the spleen from recipient mice 4 (up) or 5 (bottom) days post LM-OVA infection in the in vivo co-transfer model (n = 3). E Representative flow cytometry plots and the MFI of activated (cleaved) caspase-3 expression in the WT (Ftofl/flCD4-Cre−OT-1+) and FTO KO (Ftofl/flCD4-Cre+OT-1+) CD8+ T cells stimulated in vitro with OVA257-264 peptide (left) for 24 h (n = 3). F Representative flow cytometry plots and the MFI of activated (cleaved) caspase-3 expression in the WT (Ftofl/flCD4-Cre−) and FTO KO (Ftofl/flCD4-Cre+) CD8+ T cells stimulated in vitro with anti-CD3/CD28 antibodies for 24 h (n = 3). G The protein levels of Bcl-2 and cleaved caspase-3 were measured by Western blot in WT (Ftofl/flCD4-Cre−) and FTO KO (Ftofl/flCD4-Cre+) CD8+ T cells stimulated in vitro with anti-CD3/CD28 antibodies for 48 h. Data are representative of two or three independent experiments shown as the mean ± SD. Statistical testing is depicted as two-sided, unpaired t-tests or two-way ANOVA; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.
