Fig. 7: Deficiency of CTCF O-GlcNAcylation negatively regulates maintenance and establishment of pluripotency.

a Growth curve analysis of WT_CTCF and MUT_CTCF cells in 5 days. b Cell cycle analysis of WT_CTCF and MUT_CTCF cells in 5 days. c, d, Colony formation assay for WT_CTCF and MUT_CTCF cells in 5 days. Colonies were stained for AP activity and divided into three categories (UD uniformly undifferentiated, PD partially differentiated or mixed, and D differentiated) as indicated. e Quantitative PCR analysis of lineage markers after mutating CTCF O-GlcNAcylation for 5 days. f Quantitative PCR analysis of pluripotency markers after mutating CTCF O-GlcNAcylation for 5 days. g Morphology of WT_CTCF and MUT_CTCF cells in 5 days with (bottom) or without (top) AP staining. Scale bar was 200 μm (top) and 50 μm (bottom). This experiment was performed three times, with similar results. h Morphology of WT_CTCF and MUT_CTCF cells after EB differentiation. Scale bar was 200 μm. This experiment was performed three times, with similar results. i Quantitative PCR analysis of lineage markers during embryoid body (EB) differentiation. j Reprogramming efficiency of MUT_CTCF compared with WT_CTCF. AP staining (left) and quantitation (right) of AP positive colony numbers. O OCT4, S SOX2, K KLF4. MEFs mouse embryonic fibroblasts, iPSCs induced pluripotent stem cells. See also Supplementary Fig. 6 and Supplemental Table 2. All statistical tests were two-sided Student t-test. Significance levels were indicated by asterisks: *p < 0.05; **p < 0.01; ***p < 0.001 (not significant, denoted as n.s.). All data were presented as mean ± SD (n = 3 biologically independent samples). Source data are provided as a Source Data file.