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. Author manuscript; available in PMC: 2018 May 4.
Published in final edited form as: Cell Stem Cell. 2017 Mar 30;20(5):706–719.e7. doi: 10.1016/j.stem.2017.03.002

Figure 2. Ratio of X chromosomes to autosomes correlates with global methylation levels in pluripotent cell lines.

Figure 2

(A) Schematic of the generation of ESC/EGC-somatic hybrids.

(B) Representative ESC/MEF hybrids and EGC/MEF hybrids show activation of somatically silenced Oct4-EGFP reporter.

(C) Representative EGC/MEF and ESC/MEF hybrids show tetraploid DNA content compared to parental fusion partners using propidium iodide (PI) staining.

(D) Global methylation levels of 4n ESC-somatic and EGC-somatic hybrids and their respective 2n fusion partners as determined by RRBS analysis. One representative line for each cell type was shown. A haploid ESC (X) line was included as a control in D–E.

(E) (Top) Median global methylation levels of 4n hybrids and their respective 2n fusion partners from Figure 2D are shown. (Bottom) The number of presumed active X chromosomes (XA) for each line and the ratio of XA to autosomes (A) are shown.

(F) Methylation levels of all captured imprinted loci from ESC/EGC-somatic hybrids and their fusion partners from Figure 2D as measured by RRBS analysis.

(G) Dendrogram and heatmap representation of imprinted methylation in ESC/EGC-somatic hybrids and their somatic fusion partners. Average methylation levels across ICRs of 2 XXYY ESC/MEF hybrids, 2 XXYY EGC/MEF hybrids, 3 XXXX ESC/MEF hybrids, 3 XXXX EGC/MEF hybrids, 1 XY MEF and 1 XX MEF line are shown. XXYY, blue; XXXX, red; XX and XY MEF (black).

See also Figure S3.