Fig. 1: YTHDF2 is selectively upregulated and redistributed in early Teff and Teff-like cells.
a, b mRNA expression of m6A modifiers in human CD8 T cells activated for different time points or differentiated toward exhaustion- or memory-like stages (GSE212357) (a). mRNA expression of m6A modifiers in CD8 T cells from anti-PD-1- or cIg-treated mouse lung tumors (GSE114300) (b). c The mean fluorescence intensity (MFI) of YTHDF2 in in vitro-generated effector CD8 T cells (Teff, n = 5 independent samples) and exhausted CD8 T cells (Tex, n = 5 independent samples). d, e The MFI of YTHDF2 in splenic or tumor-infiltrating T cell subpopulations from B16F10-OVA-bearing mice at day 6 (n = 5 mice) or day 13 (n = 5 mice) after tumor inoculation. Tpex, progenitor exhausted T cells. Tmem, memory T cells. f The MFI of YTHDF2 in CD8 T cell subpopulations from anti-PD-1- or cIg-treated B16F10-OVA tumors (n = 5 mice per group). g Immunoblotting analysis of YTHDF2 in the cytosol and nucleus of stimulated mouse or human CD8 T cells. h Left panel, representative confocal immunofluorescence images of YTHDF2 (red) and DAPI (blue) in naïve or activated CD8 T cells. Scale bar, 10 μm. i Representative immunofluorescent staining of CD8 (red) and YTHDF2 (green) in progressed (n = 5 independent samples) or regressing (n = 5 independent samples) B16-OVA tumors. A dashed box represents the 4× enlarged area shown in the bottom panels with separate channels. White arrows point to cells positive for YTHDF2 and CD8. Scale bar, 10 μm. Middle panel, frequencies of YTHDF2-positive CD8 T cells. Right panel, quantification of the nuclear to cytoplasmic ratios of YTHDF2 intensity in YTHDF2-positive CD8 T cells. j Representative confocal immunofluorescence images of YTHDF2 (red) and DAPI (blue) in anti-PD-1- or cIg-treated CD8 Tpex cells (n = 6 independent samples per group). Scale bar, 10 μm. Error bars, mean ± s.e.m. One-way analysis of variance (ANOVA) (d, e) or two-tailed unpaired Student’s t-test (c, f, i, j).
