Extended Data Fig. 9: The protein levels of TE down-regulated mRNAs.
From: tRNA-m1A modification promotes T cell expansion via efficient MYC protein synthesis
a. Naïve CD4+ T cells were activated with 5 μg/mL anti-CD3 antibody and 2 μg/mL anti-CD28 antibody for 6 hours. The expression of Rhoa was verified by RNA sequencing. n = 2 biologically independent samples. NS: non-significant; two-tailed, unpaired t-test. b. Naïve CD4+ T cells obtained from the spleens of Trmt61a-KO and littermate control mice were stimulated for the indicated times with anti-CD3 and anti-CD28 antibodies (6 hours). The representative target with TE-downregulation was verified by immunoblot. Representative data of three independent experiments are shown. c. Retrovirus-mediated expression of RHOA-WT and RHOA-Mutant (All Leu codons replaced by CTT, and all Gly codons replaced by GGT) in WT and Trmt61a-KO CD4+ T cells. The protein level of RHOA was quantified by immunoblot. Representative data of two independent experiments are shown. d. Retrovirus-mediated expression of CDK2-WT and CDK2-Mutant (All Leu codons replaced by CTT, all His codons replaced by CAT, and all Gly codons replaced by GGT) in WT and Trmt61a-KO CD4+ T cells. The protein level of CDK2 was quantified by immunoblot. Representative data of two independent experiments are shown. e. Heatmap showing the protein expression of TE-down, TE-up, and TE-unchanged transcripts in Trmt61a-KO activated CD4+ T cells (6 hours), data obtained from the PRIDE database under accession numbers PXD004367. f. Venn diagram showed the intersection of the TE-down genes in RiboTag RNA-seq and genes in WPC-2 or WPC-3 (WPC-2 and WPC-3 were obtained from PMID: 28285833). The P value of intersection was calculated by the fisher test (two-sided).
