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. 2014 Feb 21;9(2):e89451. doi: 10.1371/journal.pone.0089451

Figure 6. ASB4 promotes TB cell differentiation in vitro.

Figure 6

A) TB stem cells (TBSCs) were isolated from wild-type and Asb4−/− extraembryonic ectoderm at E7.5. Cells isolated from each conceptus were cultured in isolation, and these data represent 4 unique populations of cells for each genotype. Serum withdrawal induces the formation of large, multinucleated trophoblast giant cells (TGCs, arrows) that differentiate from TBSCs (asterisks). As shown, wild-type TBSCs largely differentiate into TGCs (left panel) while Asb4−/− cells remain in undifferentiated embryoid bodies (right panel). MEF-feeder cells are indicated by filled arrows. Dashed outlines indicate the border of non-MEF cell clusters. B) JAR cells were transfected to express vector, Asb4, or Asb4 co-transfected with vector, wild-type Id2, or degradation-resistant Id2 (DR-Id2). ASB4 induced hCG secretion, and co-expression of wild-type ID2 with ASB4 did not change hCG secretion compared to ASB4 expression alone. DR-ID2 prevented dcbAMP-induced hGC section, with concentrations of hCG no different than vector/vector transfected cells. * p<0.01 compared with vector/vector.