Fig. 4

L1 expression is maintained upon TET-mediated demethylation. a Deep amplicon sequencing from oxBS-treated DNA was used to measure 5mC and 5hmC levels at young L1s in WT and TET-depleted ESCs; each data point represents the average value from three biological replicates at a given CpG within the amplicon. b Double TET1/TET2 knockdown does not lead to more pronounced effects on 5mC/5hmC than TET2 knockdown alone. c Quantitative reverse transcription polymerase chain reaction (RT-qPCR) data of TET1- and/or TET2-depleted ESCs, at four or ten days following lentiviral shRNA delivery (n = 6); no statistically significant differences are detected (t-test). d Western blot for ORF1p also shows no difference in expression at the protein level. e Representative northern blot for L1Tf, with averaged data from n = 4 quantified on the right; no statistically significant differences in the levels of full-length L1Tf are detected (t-test). f RT-qPCR (n = 6) and oxBS (n = 3) analysis of individual L1 copies reveals a similar pattern to that seen in the pool of all L1s, with no expression differences despite increased 5mC levels. * p < 0.05, *** p < 0.001, ANOVA with post-hoc Tukey test