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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: A cytokine screen using CRISPR-Cas9 knock-in reporter pig iPS cells reveals that Activin A regulates NANOG

Fig. 2

Verification and transcriptome analysis of NANOG tdTomato knock-in and WT PC-iPS cells. A NANOG tdTomato knock-in positive PC-iPS cells (nanog) could be clustered with pig EPS cells (pEPSC), but separate with trophoblast cells (TE), inner cell mass (ICM), and early blastocysts (SB). B Knock-in positive PC-iPS cells were expressed OCT4 and SOX2 pluripotent markers. Scale bar, 20 μm. C (a) knock-in positive PC-iPS cells formed EB spheres. Scale bar, 50 μm; (b) knock-in positive PC-iPS cells were differentiated to three germ layers which were ectodermal (β-TUBULIN), mesodermal (α-SMA), and endodermal (VIMENTIN). Scale bar, 200 μm. D Heat map of the clustering analysis for expression of selected TGF-β signal pathway-related genes in NANOG tdTomato knock-in versus WT PC-iPS cells. E (a) Pluripotency-related genes were up-regulated in NANOG tdTomato knock-in positive PC-iPS cells comparing with WT PC-iPS; (b) TGF-β signal pathway-related genes were also upregulated in NANOG tdTomato knock-in positive PC-iPS cells. Values were normalized as log2(FPKM+1), where FPKM is fragments per kilobase of exon per million reads mapped. F RT-PCR analysis of expression of NANOG, TCFP2L1, and CDH1 in NANOG tdTomato knock-in and WT PC-iPS cells. ****< 0.0001; ***< 0.001; **< 0.01, *< 0.5; ns, not significant

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