1. Developmental Biology
  2. Stem Cells and Regenerative Medicine
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Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

  1. Sridhar Selvaraj
  2. Ricardo Mondragon-Gonzalez
  3. Bin Xu
  4. Alessandro Magli
  5. Hyunkee Kim
  6. Jeanne Lainé
  7. James Kiley
  8. Holly Mckee
  9. Fabrizio Rinaldi
  10. Joy Aho
  11. Nacira Tabti
  12. Wei Shen
  13. Rita CR Perlingeiro  Is a corresponding author
  1. University of Minnesota, United States
  2. Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), Mexico
  3. Sorbonne Universités, Faculté de Médecine site Pitié-Salpêtrière, France
  4. Bio-Techne, United States
Research Article
Cite this article as: eLife 2019;8:e47970 doi: 10.7554/eLife.47970
8 figures, 1 table and 1 additional file

Figures

Figure 1 with 3 supplements
Combinatorial treatment with four small molecules augments myotube generation from human PS cells and their fusion ability.

(A) Bar graph shows expression profile of MYH isoforms in hiPSC-1-derived myotubes. Data are shown as mean ± S.E.M.; n = 3, ***p<0.001. (B) Bar graph shows the ratio of % MyHC-stained area to % DAPI area in myotubes resulting from treatment with five candidates identified by the small molecule screening. Data show significant increase (***p<0.001) compared to DMSO in all three PS cell lines analyzed (hESC-1, hiPSC-1 and hiPSC-2). Data from three independent replicates are shown, normalized to DMSO, as mean ± S.E.M. (C) Bar graph shows the ratio of % MyHC-stained area to % DAPI area in iPS cell-derived myotubes that had been differentiated in the presence of all candidates combined, or with individual candidates excluded from the overall combination. Data from three independent replicates are shown normalized to DMSO. Values are shown as mean ± S.E.M. ***p<0.001. (D) Representative images show immunostaining for MyHC (in red) in hiPSC-1 myotubes differentiated with combinatory treatments of small molecules or DMSO. DAPI stains nuclei (in blue). Scale bar is 100 μm. (E) Bar graph shows fusion index analysis of myotubes that were differentiated with small molecule combinations or DMSO. Data are shown as mean of three independent replicates ± S.E.M. ***p<0.001. (F) Stacked bar graph shows the frequency of number of nuclei per myotube upon differentiation with combinatory treatments or DMSO. Data are shown as mean of three independent replicates ± S.E.M. Statistical analysis compares each combination to DMSO. *p<0.05 **p<0.01 ***p<0.001.

https://doi.org/10.7554/eLife.47970.002
Figure 1—source data 1

Tocriscreen Stem Cell Toolbox compounds tested during myogenic terminal differentiation of PS cell lines.

https://doi.org/10.7554/eLife.47970.006
Figure 1—figure supplement 1
BMP and TGFβ signaling inhibition induce somite-like specification during the in vitro muscle differentiation of iPAX7 PS cells.

(A) Schematic representation of the modified EB-iPAX7 protocol, which includes incubation with LDN193189 and SB431542, BMP and TGFβ signaling inhibitors, respectively, addition of Doxycycline on day 5, and sorting of myogenic progenitors on day 12. (B) Bar graphs show expression analysis of paraxial mesoderm (MSGN1, T and TBX6) and somite (FOXC2, MEOX1, PAX3 and TCF15) genes relative to GAPDH in hESC-1 and hiPSC-1 lines at day 4 and day 6 of the EB-iPAX7 protocol (A) with or without addition of LDN193189 and SB431542 (LS). Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 ***p<0.001.

https://doi.org/10.7554/eLife.47970.003
Figure 1—figure supplement 2
Induction of somite-like stage enhances iPAX7 PS cell-derived myogenic differentiation into myotubes.

Representative images show immunostaining for MyHC (in red) in myotubes derived from four iPS cell control (1, 2, 3 and 4), one ES cell control and two patient-specific (DMD one and DMD 2) iPS cell lines differentiated under the standard EB-iPAX7 protocol (-LS) or including LDN193189 and SB431542 treatment (+LS) for somite-like specification. DAPI stains nuclei (in blue). Scale bar is 200 μm.

https://doi.org/10.7554/eLife.47970.004
Figure 1—figure supplement 3
Small molecule screening reveals compounds that enhance myogenic differentiation efficiency.

(A) Schematic representation of the small molecule screening procedure. Each well from a 96-well plate contained an individual compound from the Tocriscreen Stem cell toolbox added in the differentiation medium. (B) Representative images of MyHC (red) immunostaining in myotubes differentiated with selected candidates upon small molecule screening (A). DAPI stains blue. Scale bar is 100 μm. (C) Bar graph shows ratio of % MyHC-stained area to % DAPI area from hiPSC-1 myotubes differentiated with compounds candidates at 5, 10 or 20 μm relative to DMSO. Data are shown as mean of three independent replicates ± S.E.M. Statistical analyses showed no significant differences among concentrations for each compound. (D) Bar graphs show gene expression analysis of MYOG and MYH isoforms relative to GAPDH of hiPSC-1 myotubes differentiated with compound candidates. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01 ***p<0.001.

https://doi.org/10.7554/eLife.47970.005
Figure 2 with 3 supplements
Combinatorial treatment with S/Da/De/F enhances the maturation of PS cell-derived myotubes.

(A) Bar graphs show the expression profile of MYOG, MYOD and MYH isoforms normalized to ACTB in hiPSC-1 myotubes differentiated with small molecule combinatorial treatment or DMSO. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01. (B) Bar graphs show expression levels of MYOG, and MYH isoforms normalized to ACTB in hESC-1, hESC-2 and hiPSC-1 myotubes differentiated with combinatory treatment or DMSO. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01. (C) Western blot shows protein expression for MYH3 (e-MyHC), MYH8 (neo-MyHC) and MYH1/2 (ad-MyHC) in hiPSC1 myotubes that had been subjected to treatment with S/Da/De/F or DMSO. Human adult skeletal muscle is shown as a reference. ACTB is used as loading control. (D, E) Representative images show immunostaining for neo-MyHC (in green) (D) and Titin (in green) (E) in hiPSC-1 myotubes differentiated in the presence of DMSO or S/Da/De/F. DAPI stains nuclei (blue). Scale bars are 100 μm (D) and 20 μm (E).

https://doi.org/10.7554/eLife.47970.007
Figure 2—figure supplement 1
Combinatorial treatment promotes neo-MyHC protein expression.

(A) Representative image of F-actin (green) and neo-MyHC (red) immunostaining analyzed by confocal microscopy in hiPSC-1 myotubes differentiated with combinatorial treatment. DAPI stains nuclei. Mid Z section is shown. Scale bar is 20 μm. (B) Protein expression analysis for neo-MyHC and Desmin by western blot of hiPSC-1, hiPSC2 and hESC-1 myotubes differentiated in the presence of combinatorial treatment (S/Da/De/F) or DMSO. Actin is used as loading control. (C and D) Bar graph shows percentage of EdU (+) nuclei (C) and ratio of % MyHC-stained area to % DAPI area (D) at days 1, 3 and 5 of hiPSC-1 myotube differentiation in the presence of combinatorial treatment or DMSO. **p<0.01.

https://doi.org/10.7554/eLife.47970.008
Figure 2—figure supplement 2
Combinatorial treatment targets pathways associated with its individual components.

(A–D) Bar graphs show gene expression analysis, relative to GAPDH, of genes related with Dexamethasone (CEBPB, CEBPD and FKBO5, (A), DAPT (NOTCH2, HES1 and JAG1, (B), SB431542 (COL1A1, ID3 and SERPINE1, (C) and Forskolin (PPARGC1A, RGS2 and NR4A1, (D) targeted-pathways in hiPSC-1 myotubes differentiated in presence of combinatorial treatment or DMSO. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 ***p<0.001.

https://doi.org/10.7554/eLife.47970.009
Figure 2—figure supplement 3
Combinatorial small molecule treatment enhances the maturation of hiPS cell-derived myotubes generated under transgene-free differentiation conditions.

(A) Representative images of MyHC (in red) immunostaining of transgene-free hiPSC-1 myotubes differentiated in the presence of DMSO or S/Da/De/F. DAPI stains nuclei (blue). Scale bar is 200 μm. (B) Bar graphs show gene expression analysis of MYH isoforms and MYOG relative to ACTB of transgene-free hiPSC-1 myotubes differentiated in the presence of DMSO or S/Da/De/F. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01.

https://doi.org/10.7554/eLife.47970.010
Figure 3 with 1 supplement
Combinatorial treatment with S/Da/De/F enhances the maturation of MD patient-specific hiPS cell-derived myotubes.

(A) Representative images show immunostaining for MyHC (in red) in hiPS cell-derived myotubes from two DMD (DMD1 and DMD2), two DM1 (DM1-1 and DM1-2) and one LGMD2A patients differentiated with small molecule combinatorial treatment or DMSO. DAPI stains nuclei (blue). Scale bar is 100 μm. (B) Bar graphs show the expression profile of MYH isoforms and MYOG isoforms normalized to GAPDH in hiPS cell-derived myotubes from two DMD (DMD one and DMD 2), two DM1 (DM1-1 and DM1-2) and one LGMD2A patients differentiated with small molecule combinatorial or DMSO treatments. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01 ***p<0.001.

https://doi.org/10.7554/eLife.47970.011
Figure 3—figure supplement 1
Characterization of hiPSC-3, hiPSC-4 and hiPSC-DMD1 reprogrammed cell lines.

(A) Representative images show typical pluripotent colony morphology for each reprogrammed iPS cell line. Scale bar is 200 μm. (B) Representative images show immunostaining of NANOG, OCT3/4, SOX2 and SSEA-4 (in red) in reprogrammed iPS cell lines. DAPI stains nuclei (in blue). Scale bar is 200 μm. (C) Images show H and E staining of teratomas generated by reprogrammed iPS cell lines. (D) Cytogenetic analyses show normal karyotypes of reprogrammed iPS cell lines.

https://doi.org/10.7554/eLife.47970.012
Figure 4 with 1 supplement
Ultrastructural differences between S/Da/De/F- and DMSO-treated myotubes.

(A–D) DMSO-treated myotubes (control) are shown at different magnifications. (A) Low magnification shows a thin and elongated myotube. (B, C) Myofibrils from control myotubes display different degrees of sarcomeric organization. (B) Discernable A-I bands, M line, and winding Z-bands, (C) Incomplete banding pattern and Z-bodies. (D) High magnification shows one SR-TT junction at the periphery of the cell. The SR has been highlighted by artificial post-coloring. (E–H) S/Da/De/F treated myotubes are shown at different magnifications. (E) Large myotube with relatively well-organized myofibrils located at the periphery or in close proximity to large bundles of mitochondria; red arrows indicate SR-TT junctions. (F–H) Myotubes subjected to S/Da/De/F treatment also display myofibrils with various degrees of sarcomeric organization. (F) Well defined A-I pattern, well delineated Z bands and visible M lines. (G) Alternating A and I bands, but discontinuous Z-band. (H) Nascent sarcomere with undefined banding patterned and Z–bodies. (I) Large myotube with a great number of SR-TT junctions (red arrows). Notice the presence of numerous mitochondria. The junctions identified by J, K and L letters are enlarged below (J–L) Different examples of representative SR-TT junctions; the SR is highlighted by artificial post-coloring. Scale bars: 5 µm in A, E and I; 1 µm in B, C and F-H; 500 nm in D and J-L.

https://doi.org/10.7554/eLife.47970.013
Figure 4—figure supplement 1
Transmission electron microscopy reveals an enhanced fusion process and numerous SR-TT junctions in S/Da/De/F-treated PS cell-derived myotubes.

(A, B) Myotubes treated with DMSO shown at low (A) and high (B) magnifications. (C, D) Myotubes treated with S/Da/De/F shown at low (C) and high (D) magnifications. These microphotographs reveal ongoing fusion processes between apposed plasma membranes of two untreated (B, numbered 1,2), and five treated myotubes (D, numbered 1–5). The fusion path is highlighted by artificial post-coloring (orange). Black arrow heads in B and d1 delimit regions of full cell fusion. Red arrows show SR-TT junctions, which are abundant in treated myotubes (D), and generally scarce in the control; they were undetectable in the present untreated myotube (B). Bars = 20 µm in A, C; 2 µm in B, D; 500 nm in d1, d2.

https://doi.org/10.7554/eLife.47970.014
Figure 5 with 1 supplement
Combinatorial treatment increases chromatin accessibility at myogenic loci.

(A) Venn diagram displaying overlap between loci detected in 2-day S/Da/De/F- and DMSO-treated cells. (B) Heatmap shows changes in chromatin accessibility between DMSO- and S/Da/De/F-treated cells (three independent biological replicates). Loci were selected based on adjusted p-value<0.05 and log2FoldChange > 1. Loci overlapping to blacklist regions are included in this heatmap. (C) Chromatin accessibility at the genomic loci proximal to MYOG, MYH3 and CEBPD genes. Dashed black boxes indicate loci characterized by significant change in chromatin accessibility. Tracks represent snapshots from the IGV browser. (D) Selected enriched motifs identified at S/Da/De/F-specific peaks using MEME-ChIP. Plot below the sequence logo indicates distribution of the motifs across the regions used as input. (E) Table schematizing the results obtained by MEME-ChIP. Only selected motifs are displayed.

https://doi.org/10.7554/eLife.47970.015
Figure 5—figure supplement 1
Analysis of chromatin accessible peaks upon combinatorial treatment.

(A) Principal component analysis of DMSO- and S/Da/De/F-treated samples using sequencing depth coverage calculated at a list of loci comprising all ATAC-seq peaks detected in this experiment. (B) Chromatin accessibility at the genomic loci proximal to HES1, ID3 and MIR206-MIR133B genes. Dashed black boxes indicate loci characterized by significant change in chromatin accessibility. Tracks represent snapshots from the IGV browser.

https://doi.org/10.7554/eLife.47970.016
Figure 6 with 2 supplements
Combinatorial treatment induces expression of genes associated with structural maturation.

(A) Heatmap shows differentially expressed genes in hiPSC-1 myotubes upon combinatorial treatment compared to DMSO from three independent replicates. (B) Table shows muscle differentiation associated transcription factors and miRNAs that were upregulated in combinatorial treatment group when compared to DMSO group as revealed by IPA. (C–D) Bar graphs show the top physiological systems (C) and canonical pathways (D) associated with genes upregulated in combinatorial treatment group when compared to that of DMSO as revealed by IPA. (E–F) Bar graphs show the top biological processes (E) and cellular components (F) associated with genes upregulated upon combinatorial treatment based on gene ontology (GO) analysis. Data are plotted as –log (p-value) in C-F.

https://doi.org/10.7554/eLife.47970.017
Figure 6—source data 1

IPA of upstream regulators of the differentially expressed genes upon combinatorial treatment confirm the pathways targeted by the small molecules.

https://doi.org/10.7554/eLife.47970.020
Figure 6—source data 2

List shows the targets of transcription factors and miRNA that were found differentially expressed upon combinatorial treatment.

https://doi.org/10.7554/eLife.47970.021
Figure 6—figure supplement 1
Validation of selected genes revealed by RNA-Sequencing upon combinatorial treatment of PS cell-derived myotubes.

(A) Bar graphs show quantitative analysis of gene expression of selected genes associated with skeletal muscle maturation in hESC-1, hiPSC-1 and hiPSC-2 myotubes, which were differentiated with combinatorial treatment or DMSO. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01 ***p<0.001. (B) Bar graphs show expression levels of genes associated with embryonic myogenic identity relative to GAPDH in hiPSC-1 myotubes, which were differentiated with combinatorial treatment or DMSO. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01 ***p<0.001.

https://doi.org/10.7554/eLife.47970.018
Figure 6—figure supplement 2
Transcriptomic analysis of genes annotated to loci with increased accessibility following S/Da/De/F treatment.

(A) Differential expression analysis of genes annotated to peaks with increased chromatin accessibility upon S/Da/De/F treatment. Genes were annotated using GREAT following a two gene association and 500 kb regulatory domain. (B–C) Gene ontology classification of S/Da/De/F-specific peaks using DAVID. Bar graphs show Biological Process categories for (B) upregulated and (C) downregulated genes from panel A.

https://doi.org/10.7554/eLife.47970.019
Figure 7 with 1 supplement
Increased contractile force generation in PS cell-derived 3D muscle constructs upon combinatorial treatment.

(A–B) Representative twitch (A) and tetanic (B) force patterns at 0.5 Hz and 20 Hz, respectively, generated by hiPSC-1 3D muscle constructs differentiated with combinatorial treatment or DMSO. Bar graphs show the twitch force (A) and tetanic force (B) as mean of three independent replicates ± S.E.M. ***p<0.001. Nine twitch peaks and three tetanic measurements from three independent muscle constructs were used for analysis. (C) Bar graphs show myogenic genes expression analysis relative to ACTB in 3D muscle constructs differentiated with combinatorial treatment or DMSO (from A and B). Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01 ***p<0.001. (D) Protein expression analysis of MYH3 (e-MyHC) and MYH8 (neo-MyHC) by western blot of hiPSC-1 3D muscle constructs and 2D differentiated myotubes with combinatorial treatment or DMSO. Actin is shown as loading control.

https://doi.org/10.7554/eLife.47970.022
Figure 7—figure supplement 1
Differentiation of PS cell-derived 3D muscle constructs in the presence of S/Da/De/F enhances the expression of adult MYH isoforms.

(A) Representative images of PS cell-derived 3D muscle constructs differentiated in the presence of S/Da/De/F or DMSO. Images were taken on the day of functional assessment. Scale bar is 1 cm. (B) Bar graphs show gene expression analysis of MYH isoforms and MYOG relative to ACTB in hiPSC-1 myotubes differentiated in 2D and 3D cultures. Data are shown as mean of three independent replicates ± S.E.M. *p<0.05 **p<0.01 ***p<0.001. (C) Representative images show immunostaining for neo-MyHC, pan-MyHC and α-actinin in longitudinal cryosections of 3D muscle constructs differentiated with S/Da/De/F or DMSO. DAPI stains nuclei. Scale bar is 100 μm.

https://doi.org/10.7554/eLife.47970.023

Tables

Key resources table
Reagent type
(species) or
resource
DesignationSource or referenceIdentifiersAdditional information
Cell line (Homo sapiens, Male)hiPSC-1PMID: 22560081PLZControl line, available with the Rita Perlingeiro lab
Cell line (Homo sapiens, Male)hiPSC-2PMID: 26411904TC-1133Control line, available with RUCDR Infinite Biologics
Cell line (Homo sapiens, Male)hiPSC-3This studyMNP-120Control line, available with the Rita Perlingeiro lab
Cell line (Homo sapiens, Female)hiPSC-4This studyMNP-119Control line, available with the Rita Perlingeiro lab
Cell line (Homo sapiens, Male)hESC-1WiCellH9ESC control line (WA09)
Cell line (Homo sapiens, Female)hESC-2WiCellH1ESC control line (WA01)
Cell line (Homo sapiens, Male)hiPSC-DMD1This studyDMD1108DMDΔex31, available with the Rita Perlingeiro lab
Cell line (Homo sapiens, Male)hiPSC-DMD2PMID: 28658631DMD1705DMDΔex52-54, available with the Rita Perlingeiro lab
Cell line (Homo sapiens, Male)hiPSC-DM1-1PMID: 29898953DM1-12,000 CTG repeats in 3'UTR of DMPK gene, available with the Rita Perlingeiro lab
Cell line (Homo sapiens, Male)hiPSC-DM1-2PMID: 29898953DM1-21,500 CTG repeats in 3'UTR of DMPK gene, available with the Rita Perlingeiro lab
Cell line (Homo sapiens, Female)hiPSC-LGMD2APMID: 315010339015CAPN3Δex17-24, available with the Rita Perlingeiro lab
Chemical compound, drugTocriscreen Stem Cell ToolboxTocrisCat# 506010 µM of each compound
Chemical compound, drugCHIR99021TocrisCat# 442310 µM
Chemical compound, drugLDN193189Cayman chemicalCat# 19396200 nM
Chemical compound, drugSB431542Cayman chemicalCat# 1303110 µM
Chemical compound, drugDAPTCayman chemicalCat# 1319710 µM
Chemical compound, drugDexamethasoneCayman chemicalCat# 1101510 µM
Chemical compound, drugForskolinCayman chemicalCat# 1101810 µM
Chemical compound, drugPD0325901Cayman chemicalCat# 1303410 µM
Chemical compound, drugDoxycyclineSigma AldrichCat# D98911 µg/ml
Recombinant proteinRecombinant Human FGF-basicPeprotechCat# 100-18B5 ng/ml
Recombinant proteinRecombinant Human HGFStem Cell TechnologiesCat# 7801910 ng/ml
Recombinant proteinRecombinant Human IGF-1Stem Cell TechnologiesCat# 780222 ng/ml
Commercial assay or kitiClick EdU Andy Fluor 555 Imaging KitGeneCopoeiaCat# A004Cell proliferation assay
AntibodyMHC (all isoforms), mouse monoclonalDSHBCat# MF20, RRID: AB_2147781Dilution-1:100 (IF)
AntibodyDesmin, mouse monoclonalSCBTCat# sc-23879, RRID: AB_627416Dilution-1:500 (WB)
AntibodyACTB, mouse monoclonalSCBTCat# sc-4778, RRID: AB_626632Dilution- 1:1000 (WB)
AntibodyTitin, mouse monoclonalDSHBCat# 9D10, RRID: AB_528491Dilution- 1:50 (IF)
AntibodyMyHC-neo, mouse monoclonalDSHBCat# N3.36, RRID: AB_528380Dilution- 1:50 (IF), 1:200 (WB)
AntibodyMyHC-neo, mouse monoclonalLeicaCat# MHCN, RRID: AB_563900Dilution- 1:20 (IF), 1:200 (WB)
AntibodyMyHC-emb, mouse monoclonalDSHBCat# F1.652, RRID: AB_528358Dilution- 1:200 (WB)
AntibodyMYH1/2, mouse monoclonalDSHBCat# SC-71, RRID: AB_2147165Dilution- 1:200 (WB)
Antibodyα-actinin, mouse monoclonalThermofisherCat# MA122863, RRID: AB_557426Dilution- 1:25 (IF)
AntibodyOCT3/4, mouse monoclonalSCBTCat# C-10, RRID: AB_628051Dilution- 1:50 (IF)
AntibodySOX2, goat polyclonalSCBTCat# Y-17, RRID: AB_2286684Dilution- 1:50 (IF)
AntibodyNANOG, mouse monoclonalSCBTCat# H-2, RRID: AB_10918255Dilution- 1:50 (IF)
AntibodySSEA4, mouse monoclonalSCBTCat# sc-21704, RRID: AB_628289Dilution- 1:50 (IF)
AntibodyAnti-mouse IgG HRP-linked (sheep polyclonal)GE HealthcareCat# NA931, RRID: AB_772210Dilution- 1:20000 (WB)
AntibodyAlexa fluor 555 goat anti-mouse IgG (goat polyclonal)ThermofisherCat# A-21424, RRID: AB_141780Dilution- 1:500 (IF)
OtherAlexa Fluor 488 Phalloidin, F-actin probeThermofisherCat# A12379Dilution- 1:40 (IF)
Sequence-based reagentMYH1ThermofisherAssay ID: Hs00428600_m1Taqman probe for RT-qPCR
Sequence-based reagentMYH2ThermofisherAssay ID: Hs00430042_m1Taqman probe for RT-qPCR
Sequence-based reagentMYH3ThermofisherAssay ID: Hs01074230_m1Taqman probe for RT-qPCR
Sequence-based reagentMYH7ThermofisherAssay ID: Hs01110632_m1Taqman probe for RT-qPCR
Sequence-based reagentMYH8ThermofisherAssay ID: Hs00267293_m1Taqman probe for RT-qPCR
Sequence-based reagentMYOD1ThermofisherAssay ID: Hs02330075_g1Taqman probe for RT-qPCR
Sequence-based reagentMYOGThermofisherAssay ID: Hs01072232_m1Taqman probe for RT-qPCR
Sequence-based reagentACTBThermofisherAssay ID: Hs99999903_m1Taqman probe for RT-qPCR
Sequence-based reagentGAPDHThermofisherAssay ID: Hs99999905_m1Taqman probe for RT-qPCR
Sequence-based reagentSLNThermofisherAssay ID: Hs00161903_m1Taqman probe for RT-qPCR
Sequence-based reagentCAPN3ThermofisherAssay ID: Hs01115989_m1Taqman probe for RT-qPCR
Sequence-based reagentATP2A1ThermofisherAssay ID: Hs01115989_m1Taqman probe for RT-qPCR
Sequence-based reagentENO3ThermofisherAssay ID: Hs01093275_m1Taqman probe for RT-qPCR
Sequence-based reagentMYF6ThermofisherAssay ID: Hs00231165_m1Taqman probe for RT-qPCR
Sequence-based reagentCKMThermofisherAssay ID: Hs00176490_m1Taqman probe for RT-qPCR
Sequence-based reagentKLF4ThermofisherAssay ID: Hs01034973_g1Taqman probe for RT-qPCR
Sequence-based reagentTNNT3ThermofisherAssay ID: Hs00952980_m1Taqman probe for RT-qPCR
Sequence-based reagentCDH11ThermofisherAssay ID: Hs00901479_m1Taqman probe for RT-qPCR
Sequence-based reagentEYA2ThermofisherAssay ID: Hs00193347_m1Taqman probe for RT-qPCR
Sequence-based reagentFSTThermofisherAssay ID: Hs01121165_g1Taqman probe for RT-qPCR
Sequence-based reagentCEBPBThermofisherAssay ID: Hs00270923_s1Taqman probe for RT-qPCR
Sequence-based reagentCEBPDThermofisherAssay ID: Hs00270931_s1Taqman probe for RT-qPCR
Sequence-based reagentFKBP5ThermofisherAssay ID: Hs01561006_m1Taqman probe for RT-qPCR
Sequence-based reagentNOTCH2ThermofisherAssay ID: Hs01050702_m1Taqman probe for RT-qPCR
Sequence-based reagentHES1ThermofisherAssay ID: Hs00172878_m1Taqman probe for RT-qPCR
Sequence-based reagentJAG1ThermofisherAssay ID: Hs01070032_m1Taqman probe for RT-qPCR
Sequence-based reagentCOL1A1ThermofisherAssay ID: Hs00164004_m1Taqman probe for RT-qPCR
Sequence-based reagentID3ThermofisherAssay ID: Hs00954037_g1Taqman probe for RT-qPCR
Sequence-based reagentSERPINE1ThermofisherAssay ID: Hs00167155_m1Taqman probe for RT-qPCR
Sequence-based reagentPPARGC1AThermofisherAssay ID: Hs00173304_m1Taqman probe for RT-qPCR
Sequence-based reagentRGS2ThermofisherAssay ID: Hs01009070_g1Taqman probe for RT-qPCR
Sequence-based reagentNR4A1ThermofisherAssay ID: Hs00374226_m1Taqman probe for RT-qPCR

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