Extended Data Fig. 8: tRNA-Lys-TTT-1-1 overexpression in CRC tissues.
(a) In RKO cells, different types of histones were overexpressed, and western blot experiments confirmed the expression efficiency of these exogenous histones. (b) Northern blot analysis was used to determine the levels of tRNA-Lys-TTT-1-1 in CRC tissue compared to adjacent normal tissues. Relative expression levels were quantified by ImageJ software, with sample #1 used for normalization. The data shows that the levels of tRNA-Lys-TTT-1-1 were significantly upregulated in CRC tissue compared to adjacent normal tissues (paired dot plots, n = 6 biologically independent samples per group, CRC cohort 3, p = 0.0043, two-tailed paired t-test). (c) The actinomycin D transcription inhibition assay revealed that the degradation of tRNA-Lys-TTT-1-1 was accelerated upon silencing of TRMT6. SW480 cells with TRMT6 silencing and control cells were treated with actinomycin D for different durations and then subjected to northern blot analysis with the indicated probes (left panel); the relative levels of remaining tRNA-Lys-TTT-1-1 were quantified using ImageJ software, with actinomycin D-untreated samples used for normalization (right panel). The experiments in a and c were independently performed two times, all with similar results.
