Myogenic Differentiation from MYOGENIN-Mutated Human iPS Cells by CRISPR/Cas9

Figure 3

Myogenic differentiation in skeletal muscle cells derived from MYOGENIN-deficient hiPS clone number 28 cells. Myogenic differentiation flowchart of the time course (a). Morphological changes and mCherry fluorescent expression after treatment with Dox (b). Differentiated myogenic cells derived from hiPS cells with or without MYOG by Dox treatment for 7 days were immunostained with anti-MYOSIN HEAVY CHAIN (MYHC, green) antibody. Nuclei were stained with 4′6-diamidino-2-phenylindole (DAPI, blue). Scale bar, 100 μm (c). The ratio of DAPI-positive mono or multiple nuclei staining present in single MYHC-positive myofibers derived from wild-type or MYOG-mutated hiPS cells (d). Relative expression of transcripts for myogenic regulatory factors, MYOG (e), endogenous MYOD1 (f), and MRF4 (g), in differentiated myogenic cells treated with Dox for 5, 7, and 9 days. All error bars indicate ±SEM (). values are determined by a t-test from a two-tailed distribution. .

(a)

(b)

(c)

(d)

(e)

(f)

(g)