human embryonic kidney hek293 cells  (ATCC)

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: iScience

Article Title: Differential conservation analysis identifies residues defining constitutive internalization in beta-adrenergic receptors

doi: 10.1016/j.isci.2026.115033

Figure Lengend Snippet: B2AR V34A 1.33 and S41A 1.40 mutations alter the organization of receptor homomers (A) Schematic showing the experimental design of BRET for receptor-receptor associations. HEK 293 cells were transiently transfected with equal amounts of Rluc8-tagged receptor, either wild-type or with mutations, and increasing amounts of Venus-tagged receptor, either wild-type or with mutations. BRET saturation curves were used to determine the (B) BRET max and (C) the BRET 50. Data are the mean ± SEM; one-sample t test: ∗ p < 0.05. (D) Curve plotted using one-site specific binding saturation equation, representative curves shown, n = 6–8.

Article Snippet: HEK 293 , ATCC , CRL-1573; RRID:CVCL_0045.

Techniques: Transfection, Binding Assay

Journal: iScience

Article Title: Differential conservation analysis identifies residues defining constitutive internalization in beta-adrenergic receptors

doi: 10.1016/j.isci.2026.115033

Figure Lengend Snippet: The effect of B2AR mutations on basal and ligand-induced cAMP (A) cAMP accumulation in HEK 293 cells transiently transfected with HA-B2AR WT and mutants with isoproterenol treatment (1 pM-10 μM, 5-min). (B) cAMP production at the maximum isoproterenol dose (10 μM) and (C) basal cAMP production. Data are the mean ± SEM from n = 5–6 independent experiments. One-sample t test: ∗∗∗ p < 0.001. (D) EC 50 values for curves in A.

Article Snippet: HEK 293 , ATCC , CRL-1573; RRID:CVCL_0045.

Techniques: Transfection

Journal: iScience

Article Title: Differential conservation analysis identifies residues defining constitutive internalization in beta-adrenergic receptors

doi: 10.1016/j.isci.2026.115033

Figure Lengend Snippet: Double mutation decreases basal activity by increasing constitutive internalization (A and B) Cell surface expression of HEK 293 cells transiently transfected with HA-B2AR WT or mutant receptors. (A) Percent cells expressing receptor shown as fold change to the WT and (B) the amount of receptor at the plasma membrane shown as fold change to the WT. n = 5, one sample t test: ∗ p < 0.05, ∗∗ p < 0.01. (C) Whole-cell expression of B2AR-Rluc8 mutant receptors shown as fold change to the WT receptor, n = 6–10. One-sample t test: ∗∗ p < 0.01. (D and E) cAMP production in (D) basal and (E) isoproterenol (1 pM-10 μM, 5-min) treated HEK 293 cells transiently transfected with WT and V34A-S41A B2AR and matched for receptor expression at the plasma membrane . (F) Schematic demonstrating antibody conditions for labeling endocytic/constitutively internalized receptor pools and the total pool of receptor (endocytic + biosynthetic). (G) Confocal microscopy images of HEK 293 cells transiently transfected with HA-B2AR WT and mutant receptors, either fed live with anti-HA antibody (endocytic imaging) or incubated with anti-HA antibody post-fixation and permeabilization (endocytic + biosynthetic imaging). Representative images shown, n = 10 cells. Scale bars, 10 μm; inset = 3 μm. (H) Constitutive internalization measured in HEK 293 cells transiently transfected with HA-B2AR, HA-V34A-S41A B2AR, or HA-B1AR live labeled with anti-HA antibody at 4°C with/without 1-h incubation at 37°C. Cell surface expression measured via flow cytometry. Data shown as percentage change from surface expression in cells kept at 4°C, mean ± SEM. n = 4. One-way ANOVA with Šídák’s multiple comparisons test: ∗ p < 0.05; ∗∗ p < 0.01. See also .

Article Snippet: HEK 293 , ATCC , CRL-1573; RRID:CVCL_0045.

Techniques: Mutagenesis, Activity Assay, Expressing, Transfection, Clinical Proteomics, Membrane, Labeling, Confocal Microscopy, Imaging, Incubation, Flow Cytometry