Figure 4.

Characterization of Sorted MSCs Derived from IKKi-Treated hESCs

(A) Alkaline phosphatase staining and quantitative alkaline phosphatase activity assay after 14 days of osteogenic induction (OI).

(B) qRT-PCR results of osteogenic markers (RUNX2 and BGLAP) after 7 days of OI.

(C) ARS staining and quantification after 14 days of OI.

(D) Alcian blue staining and quantification after 21 days of chondrogenic induction.

(E) qRT-PCR results of chondrogenic markers (SOX9 and COL2a1) expression after 14 days of chondrogenic induction (CI).

(F) Oil red O staining and quantification after 21 days of adipogenic induction (AI). Scale bar indicates 200 μm.

(G) qRT-PCR results of adipogenic markers (PPAR-γ and LPL) expression after 14 days of AI.

(H) Bone formation in vivo by sorted MSCs derived from IKKi-treated H1 hESCs in six immunocompromised mice. Bar indicates 100 μm.

(I) Karyotype analysis of sorted MSCs derived from IKKi-treated H1 hESCs. For all in vitro experiments, three independent experiments were performed.