Fig. 5. Transcriptome-wide analysis of RNA stability in WT vs TKO ESCs.

a 5hmC does not seem to impact mRNA translation. Box plot showing the translation efficiency of non-modified and 5hmC-modified transcripts in WT ESCs using our hMeRIP-Seq data (n = 3) and published data³⁹ (two-tailed Wilcoxon rank-sum test, P > 0.05). b 5hmC leads to a shorter mRNA half-life. Box plot showing the difference in mRNA half-life between non-modified and 5hmC-modified transcripts in WT ESCs using our hMeRIP-Seq data (n = 3) and published data⁴⁰ (two-tailed Wilcoxon rank-sum test, P < 10⁻¹²). In the box plot, the boxes represent the interquartile range of the records, and the lines across the boxes indicate the median value of the records. The whiskers indicate the highest and lowest values among the records that are no more than 1.5 times greater than the interquartile range. c Scheme illustrating the protocol for transcription inhibition with α-amanitin in WT and TKO ESCs followed by RNA-Seq. Experiments were performed in biological duplicate. d Volcano plots show longer mRNA half-lives in Tet-depleted than WT ESCs. (P-value corrected for multi-testing < 0.05 and FC > 1.5). e Bart chart showing a greater number of unstable transcripts in WT vs TKO ESCs, as determined by RNA-Seq at 0 and 4 h post α-amanitin treatment.