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. 2019 Aug 28;27(12):2147–2157. doi: 10.1016/j.ymthe.2019.08.011

Figure 3.

Figure 3

Gene Correction of 0826 and 0989 iPSCs

(A and B) PCR shows amplification of the region spanning the knockin in genomic DNA from uncorrected (U) and corrected clones for samples 0826 (C5, C10) (A) and 0989 (C1, C3) (B). (C) Representative images show MHC (in red) staining in gene-corrected 0826 (C5, C10) and 0989 (C1, C3) iPSC-derived myotubes. DAPI stains nuclei (in blue). Scale bars: 100 μm. (D) RT-PCR analysis of gene-corrected and uncorrected 0826 and 0989 iPSC-derived myotubes. Upper panel shows amplification of full-length CAPN3 (Ex1-24). The second panel shows amplification of CAPN3 exon 1 to sv40pA sequence, which is specific to the insert. The lower panel indicates ACTB used as loading control. (E) Western blot shows CAPN3 protein expression in gene-corrected 0826 (left) and 0989 (right) iPSC-derived myotubes. (F) PCR shows amplification of the region spanning the knockin in genomic DNA from gene-corrected iPSCs post-excision of the selection cassette (cassette-free, CF). (G) Western blot shows CAPN3 protein expression in cassette-free gene-corrected 0826 and 0989 iPSC-derived myotubes.