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: Frontiers in Immunology

Article Title: Neuroinflammatory and transcriptional dynamics during SARS-CoV-2 infection in KRT18-hACE2 mouse brain

doi: 10.3389/fimmu.2026.1716597

Figure Lengend Snippet: Neuropathological changes and inflammatory mediators in K18-hACE mouse brain following SARS-CoV-2 infection. (A, B) Immunofluorescence staining of brain tissue from uninfected and SARS-CoV-2 infected mice showing colocalization of DAPI (blue, nuclear stain) with (A) Iba-1 (green) and (B) CD68 (red). Lower panels show single staining for Iba-1 or CD68. Scale bar: 50 µm. (C) TUNEL assay of brain tissue from uninfected and SARS-CoV-2 infected mice. (D) Quantification of panel C showing the percentage of TUNEL+ cells in each group. Student t-test was performed to determine the significance. **p < 0.01, ***p < 0.001).

Article Snippet: DNA fragmentation was detected on rehydrated paraffin-embedded sections using a one-step TUNEL in situ apoptosis detection kit (E-CK-A325; Elabscience), according to the manufacturer’s instructions.

Techniques: Infection, Immunofluorescence, Staining, TUNEL Assay

Journal: Frontiers in immunology

Article Title: Graphene quantum dots induce cascadic apoptosis via interaction with proteins associated with anti-oxidation after endocytosis by Trypanosoma brucei .

doi: 10.3389/fimmu.2022.1022050

Figure Lengend Snippet: FIGURE 3 Cytotoxicological effects graphene quantum dots (GQDs) on Trypanosoma brucei 12 and 24 h post-exposure. (A) Quantitative analysis of T. brucei intracellular reactive oxygen species (ROS) values based on the results in Supplementary Figure S7. The intracellular ROS levels increased with prolonged exposure and increased doses. (B) Alterations of the MMP (DYm) in T. brucei following treatment with GQDs. The levels of DYm decreased with increased doses and duration of exposure. A JC-10 fluorescent probe was applied for the detection using flow cytometry (Supplementary Figure S8). (C) Increased cytoplasmic Ca2+ concentration of T. brucei following treatment with GQDs for 12 and 24 h, demonstrated by flow cytometry. Changes in the intracytoplasmic Ca2+ concentration captured by confocal microscopy, which can be seen in Supplementary Figure S9. (D) Intracellular ATP was significantly decreased in T. brucei after exposure to GQDs. (E) Exposure to GQDs resulted in the developmental arrest of T. brucei. The proportions of cell cycle dispersion are shown in graphs, predicated in the flow cytometry data in Supplementary Figure S10. (F) Flow cytometry analysis of the increased DNA fragmentation in T. brucei after treatment with varying concentrations of GQDs for 12 and 24 h using TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) staining. T. brucei parasites that were not exposed to GQDs served controls. All values are the mean ± SD of triplicates. Student’s t-test was used to obtain p- values. *p < 0.05, **p < 0.01, ***p < 0.001 (compared to the control).

Article Snippet: The annexin V-FITC Apoptosis Detection Kit (C1062L), Cell Cycle and Apoptosis Analysis Kit (C1052), Cell Counting Kit-8 (C0038), and one-step TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) Apoptosis Assay Kit (C1088) were provided by Beyotime Biotechnology (Suzhou, China).

Techniques: Cytometry, Concentration Assay, Confocal Microscopy, Dispersion, Flow Cytometry, TUNEL Assay, Staining, Control

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: MEs-miR-146a protects H9c2 cells from OGD/R induced damage. ( A ) After 24 h of incubation, PKH26 labeled exosomes could be uptaken up by H9c2. Scale bar = 20 μm. ( B ) MEs-miR-146a decreased OGD/R induced cell apoptosis of H9c2 according to TUNEL assay. Scale bar = 50 μm. * P < 0.05, ** P < 0.01 versus the control group; & P < 0.05, && P < 0.01 versus the OGD/R group; ▲ P < 0.05 versus the OGD/R-MEs group. n = 6

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Incubation, Labeling, TUNEL Assay, Control

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: MEs-miR-146a protects NRCM cells from OGD/R induced damage. ( A ) Representative fluorescence images indicated that PKH26 labeled exosomes were taken up by NRCM cells after 24 h of incubation. Scale bar = 20 μm. ( B ) MEs-miR-146a decreased OGD/R induced cell apoptosis of NRCM according to TUNEL assay. Scale bar = 50 μm. ** P < 0.01 versus the control group; & P < 0.05, && P < 0.01 versus the OGD/R group; ▲ P < 0.05 versus the OGD/R-MEs group. n = 6

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Fluorescence, Labeling, Incubation, TUNEL Assay, Control

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: MEs-miR-146a exhibited better therapeutic efficacy in decreasing apoptotic cells and limiting inflammation than MEs. ( A - B ) Representative TUNEL staining images and quantification of apoptotic radio. ** P < 0.01 versus the Sham group; & P < 0.05 and && P < 0.01 versus the MIRI group. ( C - F ) The expression of inflammatory factors of rat serum were detected by ELISA. * P < 0.05, ** P < 0.01 versus the Sham group; & P < 0.05, && P < 0.01 versus the MIRI group; ▲ P < 0.05, ▲▲ P < 0.01 versus the MIRI-MEs-miR-146a group. n = 5

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Drug discovery, TUNEL Assay, Staining, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Nanobiotechnology

Article Title: Targeting delivery of miR-146a via IMTP modified milk exosomes exerted cardioprotective effects by inhibiting NF-κB signaling pathway after myocardial ischemia-reperfusion injury

doi: 10.1186/s12951-024-02631-0

Figure Lengend Snippet: Tail vein injection of IMTP-MEs-miR-146a significantly ameliorated myocardium apoptosis and attenuated inflammation at the early stage of MIRI. ( A ) Representative H&E images of heart tissue from each group. Scale bar = 50 μm. ( B - C ) Representative images of TUNEL staining 24 h after MIRI and quantitative analysis of apoptotic radio. * P < 0.05 and ** P < 0.01 versus the MIRI group; & P < 0.05 versus the MIRI-MEs-miR-146a group. n = 8. ( D - G ) The expression of inflammatory factors of rat serum were detected by ELISA. ∆∆ P < 0.01 versus the MIRI group; ▲▲ P < 0.01 versus the MIRI-MEs-miR-146a group. n = 8

Article Snippet: The one-step TUNEL Apoptosis Detection Kit (C1088, Green Fluorescence, Beyotime) was used to incubate for 60 min at RT, following by nuclear labeling with DAPI (H-1200-10, Vector Laboratories, Burlingame, CA, USA), both in the dark.

Techniques: Injection, TUNEL Assay, Staining, Expressing, Enzyme-linked Immunosorbent Assay