Figure 5.

MyoD-Mediated Directed Differentiation of hiPSC Clones to Myogenic Lineage

(A) Schematic of myogenic differentiation of hiPSCs achieved by lentivirus mediated MyoD overexpression. (B) Differentiated wild-type, DMD, and UTRNΔIMTR myotubes were stained with DAPI (blue) and MYHC (green). Scale bar, 200 μm (C) Efficiency of myogenic differentiation determined by the fusion index (percentage of MYHC-positive myotubes with more than 2 nuclei). We counted a total of 85, 98, and 80 myotubes of wild-type, DMD, and UTRNΔIMTR, respectively. Average from three wells (three random fields from each well) with ± SEM are shown. (D) Gene-expression analysis by qPCR of MyoD infected UTRNΔIMTR clone in tamoxifen untreated (day 0) and tamoxifen treated (day 4 and day 8) point. Expression of pluripotency marker NANOG, skeletal muscle marker MyoD1, MyoG, and endogenous MyoD1 are shown (n = 3).