Extended Data Fig. 9: CD160 regulates Fcer1g and Tnfrsf9 expression to affect the cytotoxic function of CD8+ T cells. | Nature Cell Biology

Extended Data Fig. 9: CD160 regulates Fcer1g and Tnfrsf9 expression to affect the cytotoxic function of CD8+ T cells.

From: CD160 dictates anti-PD-1 immunotherapy resistance by regulating CD8+ T cell exhaustion in colorectal cancer

Extended Data Fig. 9

a, Relative mRNA expression of Fcer1g in CD160−/+CD8+ T cells with or without anti-CD3/CD28 stimulation for 24 hours (n = 3 biological replicates). b, Relative mRNA expression of Cd160 in CD160−CD8+ T cells overexpressing Cd160 (n = 3 biological replicates). c, Relative mRNA expression of Fcer1g in CD160+CD8+ T cells after knocking down Fcer1g (n = 3 biological replicates). d, Relative mRNA expression of Fcer1g in CD160−CD8+ T cells overexpressing Fcer1g (n = 3 biological replicates). e, Relative mRNA expression of Tnfrsf9 in CD160−/+CD8+ T cells with or without anti-CD3/CD28 stimulation for 24 hours (n = 3 biological replicates). f, Relative mRNA expression of Tnfrsf9 in CD160+CD8+ T cells after knocking down Tnfrsf9 (n = 3 biological replicates). g, Relative mRNA expression of Tnfrsf9 in CD160−CD8+ T cells overexpressing Tnfrsf9 (n = 3 biological replicates). h–j, Representative flow cytometry plots (h) and frequencies of FcεR1γ+ (i) and 4-1BB+ (j) cells in CD160+ versus CD160− CD8+ T cells isolated from tumor tissues of CRC patients (n = 6 samples). k–m, Representative flow cytometry plots (k) and frequencies of FcεR1γ+ (l) and 4-1BB+ (m) cells in tumor-infiltrating CD160+ versus CD160− CD8+ T cells from MC38 tumor-bearing mice (n = 6 mice). n, Representative images of NF-κB p65 and NF-κB p52 protein expression in CD160−/+CD8+ T cells with or without anti-CD3/CD28 stimulation for 24 hours analysed by Western blot. o, Binding of HA-tagged CD160 to the Flag-tagged PI3K p85α and p110δ in HEK293T cells. p, PLA signals between PI3K and CD160 in CD160−CD8+ T cells. Scale bars: 10 μm. q, Predicted binding models, stability, and binding free energy of CD160 interactions with p110α, p110β, p110δ. r, Knockdown efficiency of p85α and p110δ siRNA was verified in CD160+CD8+ T cells. Data are shown as the mean ± s.e.m, P values were determined by unpaired two-tailed Student’s t-test (a, b, c, d, e, f, g, i, j, l and m).

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