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. 2020 Jun 25;48(14):7748–7766. doi: 10.1093/nar/gkaa529

Figure 1.

Figure 1.

J1, TKO and 3B3l mESCs achieve ground state in 2i. (A) Schematic showing cell line derivation. An expression construct for the co-factor Dnmt3l was added to Dnmt3a/3b knockout mESCs rescued by exogenous constitutive expression of Dnmt3b (3B) to create a Dnmt3a/3b knockout expressing both Dnmt3l and Dnmt3b (3B3l). Dnmt3l was also added to a to Dnmt3a/3b knockout mESCs (KOVI) to create KOVI-3l. (B) Morphology images of mESCs in indicated culture conditions. Scale bar 100 μm. (C) Expression levels of indicated Dnmt's and Prdm14. For each gene, fold change with respect to (wrt) average J1-serum probe value is represented (as log2) with SEMs. * indicates P < 0.05 (unpaired t-test). (D) Immunocytochemistry images of indicated conditions showing Nanog (green) and Esrrb (red) with DNA counterstaining (DAPI, blue) and merge. Scale bar 100 μm. (E) HPLC quantification of 5mC levels of indicated mESCs, bars represent mean ± SD of 2–3 biological replicates. (F) Bisulfite sequencing data for indicated gene promoters and cell lines, black: methylated CpG, grey: un-methylated CpG, white: missing data. Numbers represent percentage of methylated CpGs. * (black: methylated CpG, grey: un-methylated CpG, white: missing data). *P < 0.05, ** P = 0.00578, two2-tailed Mann–Whitney U-test.