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: Human Mutation

Article Title: A Novel Gain‐of‐Function GLUL Variant Is Associated With Developmental and Epileptic Encephalopathy With Enlarged Perivascular Spaces

doi: 10.1155/humu/2799390

Figure Lengend Snippet: Genetic and structural characterization of the GLUL c.522_536dup variant. (a) Pedigree and Sanger sequencing chromatograms. The sequence traces from the proband (upper panel) show a double peak starting from the duplication site, consistent with heterozygosity. (b) Superimposition of the mutant GS dimer (green) onto the WT GS structure (PDB 2QC8, blue). Black dotted lines indicate the active site. The magnified view on the right shows the mutated region, with the five duplicated amino acids (p.Ile175_His179dup) highlighted in magenta. (c) Western blot analysis of GS protein expression in HEK293 cells transfected with WT, c.1021C > T, or c.522_536dup constructs. GAPDH was used as a loading control. (d) GS enzyme activity in HEK293 cells transfected with WT, c.1021C > T, or c.522_536dup constructs, measured across a range of L‐glutamine concentrations (1, 2, 4, 16, 32, and 64 mM). The c.1021C > T variant showed markedly reduced activity at all concentrations except 64 mM. The c.522_536dup variant exhibited significantly higher activity than WT at 1 mM ( p < 0.0001), 2 mM ( p < 0.05), 32 mM ( p < 0.01), and 64 mM ( p < 0.01), with no significant difference at 4 or 16 mM. # p < 0.05; ## p < 0.01; ### p < 0.0001; ns, not statistically significant. (e) Location of the GLUL variants. The variant c.522_536dup identified in this study is indicated in red. Seven variants previously reported in AR GS deficiency are shown against a green background, and six variants associated with AD DEE are shown against a yellow background.

Article Snippet: HEK293 cells were maintained in DMEM media (Servicebio, #G4515) supplemented with 2% penicillin/Streptomycin (Gibco, #15140122), 10% fetal bovine serum (FBS) (Procell, #164250), and 4 mM glutamine (Servicebio, #GM3031) at 37 °C in 5% CO 2 .

Techniques: Variant Assay, Sequencing, Mutagenesis, Western Blot, Expressing, Transfection, Construct, Control, Activity Assay