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

Journal: International Journal of Nanomedicine

Article Title: Transferrin-Functionalized Conjugated Polymer Nanoparticles for Enhanced Photodynamic Therapy of Glioblastoma

doi: 10.2147/IJN.S592688

Figure Lengend Snippet: Integrated in silico and experimental analysis of transferrin receptor 1 (TfR1 / TFRC) expression in gliomas and representative cell lines. Box-plot summary of TFRC mRNA expression obtained from GEPIA (Gene Expression Profiling Interactive Analysis) based on tumor and normal samples from the TCGA and the GTEx databases (accessed March 2025). ( A ) Comparison of TFRC expression in high-grade gliomas (GBM) vs. low-grade glioma (LGG) (n indicated on each plot). ( B ) TFRC expression stratified by canonical GBM molecular subtype (classical, mesenchymal, neural, proneural). Boxes represent the interquartile range, horizontal lines the median, whiskers extend to 1.5×IQR, and individual data points are overlaid. Brackets with asterisks denote statistically significant pairwise differences (see Methods for statistical test). ( C ) TfR1 expression levels in GBM cell lines and HEK293 (non-tumor control). Data taken and adapted from the Human Protein Atlas. ( D ) Representative flow-cytometry histograms of surface TfR1 staining in U87MG, T98G, MO59K and HEK293 cells. Traces correspond to unstained control (red), secondary-only control (anti-mouse IgG–AF647; Orange) and specific anti-CD71 primary staining followed by anti-mouse-AF647 secondary (blue). The horizontal bracket on each histogram indicates the gate used to define TfR1-positive events. ( E ) gMFI of the CD71 signal (mean ± SD; n = biological replicates indicated in Methods), and ( F ) percentage of TfR1-positive cells. Data were normalized to appropriate controls. Statistical comparisons were performed as described in Methods (* p < 0.05 ).

Article Snippet: Moreover, analysis of mRNA expression data from the Human Protein Atlas indicates that the U87MG cell line (derived from GBM) exhibits elevated TFRC/TfR1 expression, whereas HEK293 cells (of human embryonic kidney origin) display minimal receptor expression ( ).

Techniques: In Silico, Expressing, Gene Expression, Comparison, Control, Flow Cytometry, Staining