Tumorigenic and Differentiation Potentials of Embryonic Stem Cells Depend on TGFβ Family Signaling: Lessons from Teratocarcinoma Cells Stimulated to Differentiate with Retinoic Acid

Figure 5

Stimulation of the Activin A/Nodal and BMP signaling pathways and inhibition of the MEK/ERK and PI3K/Act signaling pathways in ES R1 cells during the course of RA-induced differentiation. (a) Design of experiments on the induced differentiation of ES R1 cells with RA and exposure to Activin A, BMP4, PD98059, and LY294002. (b) ALP activity and Oct4 and Gata4 expression in differentiating ES R1 cells after exposure to RA, Activin A, BMP4, PD98059, and LY294002 on day 3 after RA and additive withdrawal. Nuclei were counterstained with DAPI. ActA: Activin A; BMP: BMP4; PD: PD98059; LY: LY294002. Scale bar = 100 μm. (c) The expression of Oct4, Nanog, and Mvh in differentiating ES R1 after exposure to RA, Activin A, BMP4, PD98059, and LY294002 on day 3 after RA and additive withdrawal. The gene expression levels of each gene in all experimental variants of differentiating ES R1 cells were calculated relative to the gene expression levels in control ES R1 cells on day 3 after RA withdrawal (ES R1 RA10 + 3) and statistically evaluated relative to the gene expression levels in control EC F9 cells on day 3 after RA withdrawal (EC F9 RA10 + 3). The data are represented as the mean ± s.d.; significant differences were estimated using ANOVA.

(a)

(b)

(c)