Journal: eLife

Article Title: Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

doi: 10.7554/eLife.47970

Figure Lengend Snippet: ( 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. 10.7554/eLife.47970.006 Figure 1—source data 1. Tocriscreen Stem Cell Toolbox compounds tested during myogenic terminal differentiation of PS cell lines.

Article Snippet: To determine whether small molecule compounds may enhance the maturation of PS cell-derived myotubes, we performed a small molecule library screening using the Tocriscreen stem cell toolbox kit (Tocris).

Techniques: Expressing, Derivative Assay, Staining, Immunostaining

Journal: eLife

Article Title: Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

doi: 10.7554/eLife.47970

Figure Lengend Snippet: ( 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.

Article Snippet: To determine whether small molecule compounds may enhance the maturation of PS cell-derived myotubes, we performed a small molecule library screening using the Tocriscreen stem cell toolbox kit (Tocris).

Techniques: Immunostaining, Staining, Expressing

Journal: eLife

Article Title: Screening identifies small molecules that enhance the maturation of human pluripotent stem cell-derived myotubes

doi: 10.7554/eLife.47970

Figure Lengend Snippet:

Article Snippet: To determine whether small molecule compounds may enhance the maturation of PS cell-derived myotubes, we performed a small molecule library screening using the Tocriscreen stem cell toolbox kit (Tocris).

Techniques: Control, Recombinant, Imaging, Proliferation Assay, Sequencing

Journal: Molecular Neurodegeneration

Article Title: BACE1 regulates expression of Clusterin in astrocytes for enhancing clearance of β-amyloid peptides

doi: 10.1186/s13024-023-00611-w

Figure Lengend Snippet: BACE1 deficiency enhances P38, ERK1/2, and cJun phosphorylation. A Western blot of BACE1-null and WT primary astrocytes lysates. Images indicate major bands for pJun, total Jun, pP38, total P38, pERK1/2, pJNK and Calnexin. B Quantification of pJun/total cJun, pP38/total p38, and pERK1/2 band intensity normalized to Calnexin ( N = 3, * p -value < 0.05, One-way ANOVA with Tukey post testing comparing between samples)

Article Snippet: Since insulin is already present in the G-5 supplemented astrocyte media, we inhibited insulin signaling by using BMS-754807 (SelleckChem), a potent IR tyrosine kinase phosphorylation inhibitor (Fig. C and D).

Techniques: Phospho-proteomics, Western Blot