Journal: Journal of Sport and Health Science

Article Title: Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle

doi: 10.1016/j.jshs.2025.101091

Figure Lengend Snippet: MiR-136-3p content in cell culture medium and uptake of extracellular miR-136-3p into cultured myotubes. MiRNA content in (A) human myotubes and (B) human pancreatic islets culture media. Results were first normalized using RNU1A1 and then presented in relation to miR-23a-3p content for n = 4 different donors for myotubes cultures and n = 4 donors for human islets. Left panel (C) shows bright-field image of cultured human myotubes and right panel (C) shows a representative fluorescence image of cultured human myotubes with cells exposed to human serum-derived EVs loaded with Cy3-miR-136-3p. Cy3 fluorescence (red) is detected in the whole cytoplasm of the human myotubes. (D) Representative fluorescence image of human myotubes exposed to HEK293 culture medium with EVs loaded with Cy3-miR-136-3p (red). (E) Representative image of human myotubes exposed to EVs loaded with TexasRed-labeled with a control RNA (orange). Nuclear Hoechst staining is shown in blue. Scale bar = 100 μm. EVs = extracellular vesicles; HEK293 = human embryonic kidney; miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.

Article Snippet: Human embryonic kidney (HEK293) cells were obtained from American Type Culture Collection (ATCC) and cultured in high-glucose (4.5 g/L) Dulbecco's Modified Eagle Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% (vol/vol) FBS.

Techniques: Cell Culture, Fluorescence, Derivative Assay, Labeling, Control, Staining

Journal: Journal of Sport and Health Science

Article Title: Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle

doi: 10.1016/j.jshs.2025.101091

Figure Lengend Snippet: NRDC is a direct target of miR-136-3p in human myotubes. Skeletal muscle NRDC mRNA is responsive to training and inactivity. (A) Tissue mRNA expression of NRDC from the Human Protein Atlas database showing enriched expression of NRDC in human skeletal muscle. (B) The miR-136-3p target site in the NRDC gene is highly conserved in mammals. (C) Luciferase activity in HEK293 cells co-transfected the NRDC 3’UTR and miR-136-3p with or without anti-miR136-3p inhibitors. miR-136-3p transfection downregulates NRDC (D) mRNA and (E) representative image of protein abundance in human myotubes. (F) Publicly available data ( GSE14413 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 6 weeks of endurance training ( n = 8). (G) Publicly available data ( GSE120862 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 2 months of aerobic training ( n = 10). (H) Publicly available data ( GSE14901 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 14 days of immobilization ( n = 24). * p < 0.05, ** p < 0.005. GSE = gene set enrichment; HEK293 = human embryonic kidney; miR = microRNA; NC = negative control; NRDC = nardilysin convertase; nTPM = normalized transcripts per million; si NRDC = small interfering RNA of NRDC ; UTR = untranslated region.

Article Snippet: Human embryonic kidney (HEK293) cells were obtained from American Type Culture Collection (ATCC) and cultured in high-glucose (4.5 g/L) Dulbecco's Modified Eagle Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% (vol/vol) FBS.

Techniques: Expressing, Luciferase, Activity Assay, Transfection, Quantitative Proteomics, Negative Control, Small Interfering RNA

Journal: Molecular Therapy Advances

Article Title: Engineered T7 RNA polymerase to improve mRNA capping efficiency and reduce dsRNA generation during in vitro transcription

doi: 10.1016/j.omta.2026.201722

Figure Lengend Snippet: mRNA generated by T7 RNAP variants had enhanced translation and reduced immunogenicity in vitro mRNA generated by the R632N and Q649L variants at 4 mM and 0.5 mM cap concentration was compared with mRNA generated by WT T7 RNAP for translation and immunogenicity in vitro . mRNAs were transfected to primary human hepatocytes to quantify (A) Cas9 expression and (B) GFP expression. HEK-Lucia RIG-I reporter cells were used to quantity immunogenicity of (C) Cas9 mRNA and (D) GFP mRNA. DC was 142-bp dsRNA as positive control and LC was lipofectamine as negative control. Cell studies were performed with three biological replicates for each condition. Data are presented as mean values, with error bars representing the standard deviation. GraphPad Prism 10.5.0 was used to perform unpaired t test to calculate significance ( p value).

Article Snippet: HEK-Lucia RIG-I reporter cells (InvivoGen) were maintained in DMEM supplemented with 10% heat-inactivated FBS (Thermo Fisher Scientific), normocin (100 μg/mL), penicillin-streptomycin (100 U/mL), blasticidin (30 μg/mL), and zeocin (100 μg/mL).

Techniques: Generated, Immunopeptidomics, In Vitro, Concentration Assay, Transfection, Expressing, Positive Control, Negative Control, Standard Deviation

Journal: Molecular Therapy Advances

Article Title: Engineered T7 RNA polymerase to improve mRNA capping efficiency and reduce dsRNA generation during in vitro transcription

doi: 10.1016/j.omta.2026.201722

Figure Lengend Snippet: mRNA generated by T7 RNAP variants had enhanced translation and reduced immunogenicity in vivo Mice were dosed intravenously with Cas9 mRNA generated by the R632N and Q649L variants at 4 and 0.5 mM cap concentration. These were compared to mRNA produced by WT T7 RNAP to assess (A) Cas9 expression, (B) IFN-α, (C) IFN-β, and (D) IP-10. HEK-Lucia RIG-I reporter cells were used to quantify immunogenicity of (C) Cas9 mRNA and (D) GFP mRNA. Phosphate-buffered saline (PBS) was used as control. Data are presented as mean, with error bars representing the standard deviation ( n = 5 mice). GraphPad Prism 10.5.0 was used to perform unpaired t test to calculate significance ( p value).

Article Snippet: HEK-Lucia RIG-I reporter cells (InvivoGen) were maintained in DMEM supplemented with 10% heat-inactivated FBS (Thermo Fisher Scientific), normocin (100 μg/mL), penicillin-streptomycin (100 U/mL), blasticidin (30 μg/mL), and zeocin (100 μg/mL).

Techniques: Generated, Immunopeptidomics, In Vivo, Concentration Assay, Produced, Expressing, Saline, Control, Standard Deviation