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. Author manuscript; available in PMC: 2018 Sep 1.
Published in final edited form as: Stem Cells. 2018 Mar;36(3):349–362. doi: 10.1002/stem.2746

Figure 2. Osteogenic differentiation of hESCs.

Figure 2

(A) Schematic of the osteogenic induction protocol.

(B) Immunostaining with an osteocalcin antibody reveals presence of bone matrix in the vicinity of black deposits. Alizarin Red S staining detected calcium ions in the same areas. Scale bars = 100 μm.

(C) Quantitative measurement of deposited calcium over time, n=5 independent replicates ± SD, *P<0.05 Student’s t test compared to d0.

(D) Quantitative mRNA analysis of RUNX2, SATB2 and OCN, normalized to GAPDH (n=3 independent samples ± SD), *P<0.05 One-Way ANOVA over day 0. Triplicates were pooled for RT-PCR analyses of the osteocyte genes CAPG and DESTRIN (inset).

OCN, osteocalcin; Aliz S, Alizarin Red S; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.