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: Life Science Alliance

Article Title: The noncoding RNA GRAS1 interacts with NKAP and protects against DNA damage and cell death

doi: 10.26508/lsa.202403105

Figure Lengend Snippet: The cell line name is indicated on the left side of each expression track.

Article Snippet: Human lung adenocarcinoma A549 cell line (ATCC No. CCL-185), human colorectal carcinoma HCT116 cell line (ATCC No. CCL-247), and human kidney embryonic HEK293 cell line (ATCC No. CRL-1573) were purchased from ATCC.

Techniques: Expressing

Journal: Life Science Alliance

Article Title: The noncoding RNA GRAS1 interacts with NKAP and protects against DNA damage and cell death

doi: 10.26508/lsa.202403105

Figure Lengend Snippet: The cell line and replicate are indicated on the left side of each expression track.

Article Snippet: Human lung adenocarcinoma A549 cell line (ATCC No. CCL-185), human colorectal carcinoma HCT116 cell line (ATCC No. CCL-247), and human kidney embryonic HEK293 cell line (ATCC No. CRL-1573) were purchased from ATCC.

Techniques: Expressing

Journal: Life Science Alliance

Article Title: The noncoding RNA GRAS1 interacts with NKAP and protects against DNA damage and cell death

doi: 10.26508/lsa.202403105

Figure Lengend Snippet: (A) CCK8 assay for survival of A549, HCT116, and HEK293 cells transfected with control GapmeR compared with GRAS1 GapmeR1 or GRAS1 GapmeR2; N = 3. (B) CCK8 assay for survival of A549 and HCT116 cells transfected with empty vector compared with SNORA13 vector; N = 4. (C) CCK8 assay for survival of A549 and HCT116 cells transfected with empty vector compared with TIGA1 vector; N = 4. (D) Schematic gene structure of GRAS1 GapmeR–resistant construct. (E) CCK8 assay for survival of A549 and HCT116 GRAS1 knockdown cells complemented with GRAS1 GapmeR–resistant vector compared with empty vector; N = 4. (F) CCK8 assay for survival of A549 and HCT116 GRAS1 knockdown cells complemented with SNORA13 vector compared with empty vector; N = 4. (G) CCK8 assay for survival of A549 and HCT116 GRAS1 knockdown cells complemented with TIGA1 vector compared with empty vector; N = 4. Error bars represent SD in all panels; (ns) P > 0.05; (*) P ≤ 0.05; (**) P ≤ 0.01; (***) P < 0.001 by two-tailed t test.

Article Snippet: Human lung adenocarcinoma A549 cell line (ATCC No. CCL-185), human colorectal carcinoma HCT116 cell line (ATCC No. CCL-247), and human kidney embryonic HEK293 cell line (ATCC No. CRL-1573) were purchased from ATCC.

Techniques: CCK-8 Assay, Transfection, Control, Plasmid Preparation, Construct, Knockdown, Two Tailed Test