Figure 5.

Rescue of DGCR8 Restored Differentiation Potential of Dgcr8^Δ/Δ iPSCs

(A) Immunoblot of DGCR8 (top) and GAPDH (bottom) in wild-type ESC, Dgcr8^Δ/Δ TTF-iPSC, and DGCR8-rescued Dgcr8^Δ/Δ TTF-iPSC extracts.

(B) Cell-cycle analyses of Dgcr8^Δ/Δ iPSCs and rescued Dgcr8^Δ/Δ iPSCs. n = 3 independent biological repeats.

(C) Colony-forming assay of wild-type, Dgcr8^Δ/Δ, and DGCR8-rescued Dgcr8^Δ/Δ iPSCs. Cells were first induced to differentiate by treatment with retinoic acid for the indicated days and then returned to conditions permissive to self-renewal for 7 days. Colonies positive for AP were scored. n = 3 independent biological repeats. Error bar, SD. ^∗p < 0.05; ^∗∗p < 0.01; Student’s t test between Dgcr8^Δ/Δ and rescued iPSCs.

(D–E″) Teratoma analyses. Shown are teratomas generated by (D) Dgcr8^Δ/Δ iPSCs, which contain virtually no differentiated somatic tissues and (E–E″) the DGCR8-rescued Dgcr8^Δ/Δ iPSCs, which contain tissues from all three embryonic germ layers: (E) neural epithelium, (E′) cartilage and muscle, and (E″) respiratory epithelium. Scale bar, 100 μm.