Supplementary Figure 1: 5hmC and 5mC analysis of TKO hESCs.

a, TET expression in WT hESCs by RNA-seq; n = 2 independent experiments. Data are presented as means ± s.d. b, Analysis of 5hmC levels in WT and knockout hESCs by 5hmC dot blot. Human fibroblasts (Fib CTRL) are used as a negative control. c, Analysis of 5hmC (left) and 5mC (right) levels in WT and TKO MEL-1 hESCs by mass spectrometry. For WT cells, two different lines were used for mass spectrometry measurements; for TKO cells, two different passages of the same line were used for mass spectrometry measurements. Human fibroblasts were used as a negative control for mass spectrometry analysis of 5hmC. Data are presented as means ± s.d. Black lines indicate comparisons to WT. Statistical analysis was performed by one-way ANOVA: *P < 0.05. d, FACS analysis for pluripotency-associated cell-surface markers TRA-1-60 and TRA-1-81 in WT and TKO hESCs; the fluorescence intensity (left) and percentage positive cells (right) are shown; n = 3 independent experiments. Data are presented as means ± s.d. Statistical analysis was performed by Student’s t test (two-sided). e, Methylation levels at HOXA7 and HOXA9 promoters in WT and TKO hESCs by WGBS; n = 1 independent experiment. The x-axis numbers correspond to the positions of CpG sites in the hg19 genome assembly