tocriscreen stem cell library  (Tocris)

Journal: eLife

Article Title: A multiplexed, single-cell sequencing screen identifies compounds that increase neurogenic reprogramming of murine Muller glia

doi: 10.7554/eLife.92091

Figure Lengend Snippet: ( A ) Bar plot representing the distribution of targets for the 92 compounds included in the screen. ( B ) Schematic of the experimental design for the small molecule screen. ( C ) UMAP of the MG and MG-derived cell types. Cells are colored by cell type. ( D ) UMAP from C with cells colored by collection. ( E ) Dot plot of the genes used to define the MG-derived cell types from the screen. Dot size indicates the percent of cells that express the gene of interest. The color indicates the log10 mean UMIs per cell. ( F ) Quantification of the fold change in Neuron cell counts between each indicated condition and the Ascl1 only control. The plots from Collections 1 and 2 used only the control wells collected on their respective days. The dose of each compound is indicated by the size of the dot. ( G ) Quantification of the total cell counts for each treatment. The compound’s dose is indicated by the size of the dot. ( H ) Plots displaying the fold change of Neuron (red) and ProL (orange) cell counts compared to Ascl1 only across all doses for the top hits from the screen. All conditions in which at least 20 cells were recovered are displayed. Figure 2B is created with BioRender.com . Figure 2—source data 1. Small molecules used from Tocriscreen Stem Cell Library. Figure 2—source data 2. Genes used and GO annotations in unknown clusters.

Article Snippet: Drugs used were all from the Tocriscreen Stem Cell Library (Tocris, 7340) and were administered in 1 μl of dimethyl sulfoxide (DMSO) at 10 mM.

Techniques: Derivative Assay, Control

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: 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: Chemical compound, drug , Tocriscreen Stem Cell Toolbox , Tocris , Cat# 5060 , 10 µM of each compound.

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: Chemical compound, drug , Tocriscreen Stem Cell Toolbox , Tocris , Cat# 5060 , 10 µM of each compound.

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: Chemical compound, drug , Tocriscreen Stem Cell Toolbox , Tocris , Cat# 5060 , 10 µM of each compound.

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