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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Cell Stem Cell. 2021 Jan 14;28(4):748–763.e7. doi: 10.1016/j.stem.2020.12.009

Figure 1. Key crotonyl-CoA-producing enzymes are induced and enriched in endodermal cells.

Figure 1.

(A) Key metabolic reactions that produce crotonyl-CoA. ACSS2, acyl-CoA synthetase short chain family member 2; ACADS, acyl-CoA dehydrogenase short chain; ACOX1, acyl-CoA oxidase 1; ACOX3, acyl-CoA oxidase 3; ECHS1, enoyl-CoA hydratase, short chain 1; GCDH, glutaryl-CoA dehydrogenase.

(B) ACSS2, ACADS, and ACOX3 are induced during endodermal differentiation of hESCs in vitro. The mRNA levels of the indicated genes in mel1 hESC-differentiated endoderm, mesoderm, or ectoderm were analyzed by qPCR (n = 3 biological replicates/group, *p < 0.05, values are expressed as mean ± SEM).

(C) ACSS2 is enriched in endodermal cells in mouse E7.5 embryos. Expression of ACSS2 was analyzed by immunofluorescence staining in E7.5 mouse embryos. The full embryo images were taken by tiling/stitching multiple fields using ZEN software. Representative images from at least three different embryos are shown. Scale bar, 100 μm.

(D) ACSS2, ACADS, and ACOX3 are induced and translocated into the nucleus during endodermal differentiation of mel1 hESCs in vitro. Mel1 hESCs were induced to differentiate into endoderm for 3 days, and then expression of the indicated proteins was analyzed by immunofluorescence staining. Scale bars for the left 8 panels: 100 μm in the FOXA2 images and 25 μm in the ACSS2, ACADS, and ACOX3 images. Scale bars for the last panel: 25 μm in the FOXA2 image and 5 μm in the ACSS2, ACADS, and ACOX3 images.

See also Figures S1 and S2.