Figure 4.

Foxj3 and Zbtb18 prevent ESCs from differentiating into neural cells. (a, b) ESCs were pretreated with miR-219 mimics for 24 h and followed by transfection of pCMV-Foxj3 or pCMV-Zbtb18 as indicated. After 48 h, the relative level of Nestin was detected through qPCR (a) and western blot (b). Actin served as the loading control. (c, d) ESCs were pretreated with miR-219 mimics for 24 h and followed by transfection of Foxj3-siRNA or Zbtb18-siRNA. After 48 h, the relative level of Nestin was detected through qPCR (c) and western blot (d). (e) Foxj3/Zbtb18-OE ESCs were induced by RA for 10 days. The morphology was observed and photographed. Scale bars: 100 μm. (f) Control ESCs or Foxj3/Zbtb18-OE ESCs were pretreated with RA for 48 h and injected into nude mice to generate teratomas. The differentiations of the three germ layers were analyzed through H&E staining. Scale bars: 100 μm. **P<0.01; NS, no significance