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. 2020 Feb 6;14(2):241–255. doi: 10.1016/j.stemcr.2020.01.004

Figure 4.

Figure 4

The Role of Act A or BMP4 in Maintaining esASCs Developmental Potency

(A) Morphology of 2i/L-ESCs were switched to AC/L or BC/L medium at passage 18 (p18). Here, we use 2i/L-ESCs with GOF/GFP reporter. Scale bars, 200 μm.

(B) Cell proliferation curves in ESCs cultured in five different mediums. Error bars indicate SEM (n = 3). Results were obtained from three independent experiments. p values were calculated by two-way ANOVA, p < 0.05.

(C) Heatmap showing scaled expression values of a total of 2,105 DEGs (mean log2(normalized read counts) > 2, log2(fold change) > 2, adjusted p value < 0.05) in AC/L-ESCs, BC/L-ESCs, and esASCs. Significantly enriched GO terms and representative genes in each cluster are listed on the right.

(D) Single AC/L-ESCs or BC/L-ESCs (GOF/GFP) were introduced into eight-cell embryos and the GOF/GFP+ embryos were counted after 24-h in vitro culture. Error bars indicate SEM (n = 3). Results were obtained from three independent experiments. Significance was tested using the two-tailed unpaired Student's t test, p < 0.05.

(E) Chimeras (E10.5) generated with AC/L-ESCs (single cell) or BC/L-ESCs (single cell). The images show that ESCs carried tdTomato+/GOF+ reporter. Scale bars, 1 mm.

(F) Summary of E10.5 chimera assays by ESCs injection.