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. Author manuscript; available in PMC: 2013 Aug 3.
Published in final edited form as: Cell Stem Cell. 2012 Jun 28;11(2):253–263. doi: 10.1016/j.stem.2012.04.026

Figure 3. Corrected HD-iPSC NSCs Have a Phenotype Similar to That of Normal iPSC NSCs.

Figure 3

(A and B) TUNEL staining and quantification of NSCs derived from uncorrected HD-iPSCs (HD-NSCs), two lines of corrected HD-iPSCs (C116-NSCs and C127-NSCs), and healthy iPSCs (normal NSCs).

(C) NSCs derived from uncorrected HD-iPSCs showed an elevated caspase-3/7 activity upon growth factor withdrawl. +GF, normal culture condition with growth factors; −GF, growth factor deprived condition. We did not detect phenotypes in iPSCs; see Figure S3.

(D) NSCs from corrected and uncorrected HD-iPSCs show that BDNF levels in corrected NSCs are consistent with those of a normal phenotype.

(E) NSCs derived from corrected cells show higher maximum respiration capacity than NSCs from uncorrected HD-iPSCs. Cells were analyzed on a Seahorse XF24 Extracellular Flux Analyzer for oxygen consumption rate (OCR). The average OCR after addition of FCCP (an uncoupler) reflects mitochondria maximum respiration capacity. Corrected NSCs and normal NSCs showed higher maximum respiration capacity than uncorrected HD-NSCs. Also see Figure S3D.

(F) RT-qPCR of TGF-β1 showed a decreased level of TGF-β1 mRNA in HD-NSCs compared with that in corrected and normal NSCs.

(G and H) RT-qPCR and western blotting of N-cadherin showed a decreased level of N-cadherin mRNA and protein in HD-NSCs compared with that in corrected and normal NSCs.

One-way ANOVA is used for statistical analysis (*p < 0.05, **p < 0.01, ***p < 0.001).