Extended Data Fig. 8: Asparagine deprivation affects glutamine metabolism to enhance CD8 T cell activation.
(A) Lysed cells from a control medium or an asparagine-deprived medium 3-day cultured CD8 T cell. The cells were subsequently analysed using liquid chromatography–mass spectrometry (LC–MS) to assess the glutamine and glutamate (n = 3). (B) GSEA tracing for mitochondrial biogenesis in Asn-free and control cultured CD8 + T cell. ES, enrichment score; NES, normalized enrichment score; FDR, false discovery ratio. (C) The level of indicated mRNAs in control T cells and asparagine-free T cell were determined by qPCR (n = 3). (D) Naïve CD8 + T cells were cultured in CD8 differentiation conditions with either asparagine-free or control medium for three days. For the Asn-free to DF condition (Asparagine and Glutamine-free), after two days of culture in an asparagine-free medium, the cells were transferred to a medium lacking both asparagine and glutamine for one day. CD8 + T cells stained for IFN-γ and granzyme B and analysed by flow cytometry (n = 3). All data represent mean ± SEM from three independent biological replicates. One-tailed unpaired Student’s t-test was used for individual comparisons in (A, C), while one-way ANOVA with Tukey’s multiple comparisons test was applied in (D). P < 0.05 was considered statistically significant.
