Additional file 1 of Pluripotent stem cell-derived skeletal muscle fibers preferentially express myosin heavy-chain isoforms associated with slow and oxidative muscles
Creators: name: Incitti, Tania
name: Magli, Alessandro
name: Jenkins, Asher
name: Lin, Karena
name: Yamamoto, Ami
name: Perlingeiro, Rita C. R.
Publisher: name: figshare
Year: 2020
Subjects: subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Physiology
subject: Pharmacology
subject: 69999 Biological Sciences not elsewhere classified
subject: Developmental Biology
subject: 110309 Infectious Diseases
subject: FOS: Health sciences
subject: FOS: Health sciences

description: Additional file 1: Figure S1. Fiber type composition of myotubes resulting from the in vitro differentiation of PSC-derived iPax3/iPax7 myogenic progenitors and adult satellite cells. Representative images show immunofluorescence staining for pan, embryonic, type I and IIA MyHC isoforms (red) in myotubes resulting from the in vitro differentiation of iPax3 and iPax7 PSC-derived myogenic progenitors, as well as adult satellite cells (adult). Nuclei were counterstained with DAPI (blue). Magnification bar: 100 μm. Figure S2. Characterization of fiber type composition. a) Graph shows the ratio of oxidative DYS+MyHC+ type I and type IIA over glycolytic type IIX and IIB myofibers following the transplantation with iPax3 and iPax7 myogenic progenitors, and adult satellite cells. Data are shown as mean ± SEM (n = 7-8 per group). **p &lt; 0.01. b) Graph bars show percentage of DYS+MyHC+ double positive with respect to total DYS+ in TA muscles transplanted with satellite cells isolated from TA or soleus muscles. Data are shown as mean ± SEM (n = 4 per group) and no statistically significant differences were observed among groups. c) Representative images show staining for embryonic MyHC (red) and dystrophin (green) in TA muscles from NSGmdx4cv mice that had been injected with iPax3 or iPax7 myogenic progenitors. Adult satellite cells served as control. Magnification bar: 100 μm. d) Graph shows respective quantification (panel c), as indicated by the number of DYS+MyHC+ double positive with respect to total DYS+. Data are shown as mean ± SEM (n = 6-8 per group). *p &lt; 0.05 and ***p &lt; 0.001. d) Quantification of CSA (μm2) of grafts from indicated groups. Data are shown as mean ± SEM (n = 4-5 per group). *p &lt; 0.05. Figure S3. Characterization of fiber type composition following the transplantation of embryonic, fetal, and neonatal primary cells. a) Representative images show staining for MyHC isoforms (red) and dystrophin (green). Nuclei were counterstained with DAPI (blue). Magnification bar: 100 μm. b) Percentage quantification of DYS+MyHC+ double positive with respect to total donor-derived DYS+ myofibers. Data are shown as mean ± SEM (n = 4-5 per group) **p &lt; 0.01. Figure S4. Characterization of fiber type composition based on recipient genetic background. a) Representative images show staining for type I MyHC (red) and Laminin (green) in TA muscles from dystrophic NSGmdx4cv and NSG mice. b) Graph shows respective quantification (panel a), as indicated by the number of MyHC+ myofibers in TA muscles from dystrophic NSGmdx4cv and NSG mice. Data are shown as mean ± SEM (n = 6 per group). *p &lt; 0.05, **p &lt; 0.01, and ***p &lt; 0.001. c) Graph bar shows percentage of MyHC+ donor-derived myofibers from Fig. 2a in comparison to iPax3 and iPax7 samples from Fig. 1b. Data are shown as mean ± SEM (n = 6-8 per group), and no statistically significant differences were observed among groups. Figure S5. Characterization of fiber types from secondary grafts. a) Representative image shows staining for embryonic MyHC (red) and dystrophin (green) in TA muscles of secondary recipients that had been injected with iPax3-derived MNCs. Magnification bar: 100 μm. b) Graph shows respective quantification (panel a), as indicated by the number of DYS+MyHC+ double positive with respect to total DYS+ myofibers of secondary grafts (iPax3 MNCs) in comparison to primary-injected TAs (iPax3). Data are shown as mean ± SEM (n = 4 per group). **p &lt; 0.01.