Fig. 4
Mutant mitochondrial tRNATrp strongly impairs neuronal maturation, but does not affect skeletal muscle development. a Experimental design used to identify patient-specific disease phenotypes triggered by m.5541C > T. b Representative images of myotubes after 2 weeks of differentiation; ACTA1 (red). Cell nuclei were co-stained with Hoechst 33342 (blue). c Differentiation efficiency of myoblasts into myotubes. The number of ACTA1-positive myotubes was counted using a cytometer to estimate differentiation efficiency. Assays were performed using 3 experimental replicates for both control and the patient. Error bars indicate as the means with SD. Statistical significance was evaluated by unpaired, two-tailed t-test. NS: Not Significant. d Representative images of iPSC-derived NSCs; Nestin (green). e Expression of representative NSC marker genes in iPSC-derived NSCs. f Representative images of CNS neurons after 2 weeks of differentiation; TUJ1 (red). Cell nuclei were co-stained with Hoechst 33342 (blue). g Differentiation efficiency of NSCs into CNS neurons. The number of TUJ1-positive CNS neurons was counted using a cytometer to estimate differentiation efficiency. Assays were performed using 3 experimental replicates for both control and the patient. Error bars indicate as the means with SD. Statistical significance was evaluated by unpaired, two-tailed t-test. *: P < 0.05. h Representative images of iPSC-derived NCCs; HNK1 (green). i Expression of representative NCC marker genes in iPSC-derived NCCs. j Representative images of PNS neurons after 2 weeks of differentiation; Peripherin (red). Cell nuclei were co-stained with Hoechst 33342 (blue). k Differentiation efficiency of NCCs into PNS neurons. The number of Peripherin-positive PNS neurons was counted using a cytometer to estimate differentiation efficiency. Assays were performed using 3 experimental replicates for both control and the patient. Error bars indicate as the means with SD. Statistical significance was evaluated by unpaired, two-tailed t-test. *: P < 0.05