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. 2022 Dec 20;4(1):100364. doi: 10.1016/j.xinn.2022.100364

Figure 5.

Figure 5

Perturbation experiment of M-DNB factors in DE differentiation system

(A) Experimental flow of M-DNB factors’ perturbation and hESCs’ differentiation. Reference is the process of hESCs’ differentiation, and we obtained ME cells at 24 h, DE cells at 72 h of hESCs’ differentiation. Perturbation of M-DNB factors’ expression consists of two parts. In tipping point 1, we knock down FOS and overexpress HSF1 and MYCN, respectively, at 12 h (nearly) and obtained ES-FOS-KO cells, ES-HSF1-OE cells, and ES-MYCN-OE cells at 72 h of differentiation. In tipping point 2, we overexpress MYCN, respectively, at 36 h (nearly) and obtained ME-MYCN-OE cells at 72 h of differentiation.

(B) RNA expression levels of ES and ME stage marker genes.

(C) RNA expression levels of ME and DE stage marker genes. OCT4, and SOX2 as ES marker genes; EOMES, MIXL1, and GSC as ME markers; SOX17 as DE marker genes.