Fig. 4

Dental epithelium-like cells were obtained with a high concentration of BMP4. a EB was cultured in EB medium for 8 days (day 8). b OE was cultured in EB medium for 2 days. c OE was stimulated with 30 μM BMP4 for 2 days. Scale bar, 300 μm. qRT-PCR expression levels of Oct4 (d), Pitx2 (e), Dlx2 (f), and AMBN (g). *P < 0.05 vs. OE; **P < 0.01 vs. OE; ^##P < 0.01 vs. EB. h, i Immunostaining of day 8 cells stimulated with 30 μM BMP4: DAPI (blue), Dlx2 (green), and AMBN (red). Scale bar, 50 μm. The DE-like cells expressed β-catenin (j) and p-Smad1/5/8 (k). DAPI (blue), β-catenin (red), p-Smad1/5/8 (green). Scale bar, 30 μm