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human embryonic kidney hek293 cells  (ATCC)


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    ATCC human embryonic kidney hek293 cells
    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 <t>HEK293</t> 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 = <t>human</t> <t>embryonic</t> <t>kidney;</t> miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.
    Human Embryonic Kidney Hek293 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 151 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/hek293+cells/Human+embryonic+kidney+cells%2C+293-hY2-10/pmc12811471-83-0-8
    Average 96 stars, based on 151 article reviews
    human embryonic kidney hek293 cells - by Bioz Stars, 2026-09
    96/100 stars

    Images

    1) Product Images from "Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle"

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

    Journal: Journal of Sport and Health Science

    doi: 10.1016/j.jshs.2025.101091

    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.
    Figure Legend 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.

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

    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.
    Figure Legend 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.

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

    Related Articles

    Incubation:

    Article Title: A natural depsipeptide antibiotic binds the E-site of the bacterial ribosome.
    Article Snippet: .. HEK293 cells (ATCC CRL-1573; generation 6) were seeded at 7,500 cells per well and HepG2 cells (ATCC HB8065; generation 18) were seeded at 4,000 cells per well in 384-well tissue culture-treated white plates in 50 μl DMEM and incubated for 18 h at 37 °C in the presence of 5% CO2. .. After 18 h of incubation, 500 nl of the compound and DMSO (1% final concentration) were added to the cells using a Labcyte Echo acoustic dispenser (Beckman Coulter).

    Cell Culture:

    Article Title: Genome-wide association mapping and targeted loss of function studies identify Shroom3 as a driver of hyperpolyploidy and ventricular dilation.
    Article Snippet: .. HEK293 cells (passage 7 to 9, ATCC, CRL- 1573) were cultured to 70% confluency in 100- mm plates at which time cells were transfected with 18 μg of flag-tagged plasmid DNA for each Shroom3 variant using Transporter 5 reagent (Polysciences, 26008) for 72 h according to the manufacturer’s protocol. ..

    Article Title: Transferrin receptor 1 shedding by the pro-inflammatory iRhom-ADAM17 complex and ADAM10 regulates cellular iron uptake and ferroptosis.
    Article Snippet: .. HEK293 cells (from the American Type Culture Collection, CRL-1573, RRID: CVCL-0045) and iRhom2 KO, iRhom1/iRhom2 double KO (dKO) as well as ADAM17 and/or ADAM10 KO cells24 were cultured in a humidified incubator at 37 °C with 5% CO2 in DMEM 10%, unless otherwise stated. .. DMEM 10% consists of DMEM high-glucose (Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS) (PanBiotech), 60 mg/l penicillin (SigmaAldrich) and 100mg/l streptomycin (Sigma-Aldrich).

    Transfection:

    Article Title: Genome-wide association mapping and targeted loss of function studies identify Shroom3 as a driver of hyperpolyploidy and ventricular dilation.
    Article Snippet: .. HEK293 cells (passage 7 to 9, ATCC, CRL- 1573) were cultured to 70% confluency in 100- mm plates at which time cells were transfected with 18 μg of flag-tagged plasmid DNA for each Shroom3 variant using Transporter 5 reagent (Polysciences, 26008) for 72 h according to the manufacturer’s protocol. ..

    Article Title: Structural characterization of kappa-opioid receptor dimer in complex with two G proteins.
    Article Snippet: .. HEK293 cells (ATCC, CRL-1573, lot: 3449904) were transfected with κOR with complementary components of Venus fluorescent protein (VN or VC) fused in the C-terminus for BiFC assay as described previously56. ..

    Article Title: Ubiquitin ligase ITCH regulates life cycle of SARS-CoV-2 virus
    Article Snippet: .. HEK293 cells (ATCC, CRL-3216) at approximately 40–60% confluence were transfected with mixtures of plasmid DNA and cationic polymer linear polyethylenimine (593215, Polysciences). .. Vero and vT2 cells at approximately 40–60% confluence were transfected with mixtures of plasmid DNA and jetPRIME transfection reagent (101000046, Polyplus).

    Article Title: Investigating the molecular basis of the serological Rh D-negative phenotype in Indonesia: nature, frequency, and impact for diagnostics.
    Article Snippet: .. Minigene splicing assay (MSA) was carried out to investigate the effect of candidate SNVs on splicing as previously described.19–21 Briefly, short genomic regions consisting of RHD exon 3 (for c.486+5G>A) and 5 (for c.635-1G>A), as well as the flanking intronic regions, were PCR-amplified by using the gDNAs of interest as templates and were inserted into the minigene pSP3 vector by homologous recombination.20 After confirmation by sequencing, the wild-type and mutated recombinant vectors were transfected separately into HEK293 cells (ATCC Number: CRL-1573) and the fluorescent RT-PCR profiles expressed from the minigene constructs were analyzed by capillary electrophoresis as previously described.20 A ρSNV score was calculated to assess the direct effect of the variant on exon inclusion compared to the wild-type conditions (ρSNV = 1, no effect; ρSNV < 1, decrease in full-length exon inclusion; ρSNV = 0, no full-length exon inclusion).21 ..

    Plasmid Preparation:

    Article Title: Genome-wide association mapping and targeted loss of function studies identify Shroom3 as a driver of hyperpolyploidy and ventricular dilation.
    Article Snippet: .. HEK293 cells (passage 7 to 9, ATCC, CRL- 1573) were cultured to 70% confluency in 100- mm plates at which time cells were transfected with 18 μg of flag-tagged plasmid DNA for each Shroom3 variant using Transporter 5 reagent (Polysciences, 26008) for 72 h according to the manufacturer’s protocol. ..

    Article Title: Ubiquitin ligase ITCH regulates life cycle of SARS-CoV-2 virus
    Article Snippet: .. HEK293 cells (ATCC, CRL-3216) at approximately 40–60% confluence were transfected with mixtures of plasmid DNA and cationic polymer linear polyethylenimine (593215, Polysciences). .. Vero and vT2 cells at approximately 40–60% confluence were transfected with mixtures of plasmid DNA and jetPRIME transfection reagent (101000046, Polyplus).

    Variant Assay:

    Article Title: Genome-wide association mapping and targeted loss of function studies identify Shroom3 as a driver of hyperpolyploidy and ventricular dilation.
    Article Snippet: .. HEK293 cells (passage 7 to 9, ATCC, CRL- 1573) were cultured to 70% confluency in 100- mm plates at which time cells were transfected with 18 μg of flag-tagged plasmid DNA for each Shroom3 variant using Transporter 5 reagent (Polysciences, 26008) for 72 h according to the manufacturer’s protocol. ..

    Article Title: Investigating the molecular basis of the serological Rh D-negative phenotype in Indonesia: nature, frequency, and impact for diagnostics.
    Article Snippet: .. Minigene splicing assay (MSA) was carried out to investigate the effect of candidate SNVs on splicing as previously described.19–21 Briefly, short genomic regions consisting of RHD exon 3 (for c.486+5G>A) and 5 (for c.635-1G>A), as well as the flanking intronic regions, were PCR-amplified by using the gDNAs of interest as templates and were inserted into the minigene pSP3 vector by homologous recombination.20 After confirmation by sequencing, the wild-type and mutated recombinant vectors were transfected separately into HEK293 cells (ATCC Number: CRL-1573) and the fluorescent RT-PCR profiles expressed from the minigene constructs were analyzed by capillary electrophoresis as previously described.20 A ρSNV score was calculated to assess the direct effect of the variant on exon inclusion compared to the wild-type conditions (ρSNV = 1, no effect; ρSNV < 1, decrease in full-length exon inclusion; ρSNV = 0, no full-length exon inclusion).21 ..

    Bimolecular Fluorescence Complementation Assay:

    Article Title: Structural characterization of kappa-opioid receptor dimer in complex with two G proteins.
    Article Snippet: .. HEK293 cells (ATCC, CRL-1573, lot: 3449904) were transfected with κOR with complementary components of Venus fluorescent protein (VN or VC) fused in the C-terminus for BiFC assay as described previously56. ..

    Polymer:

    Article Title: Ubiquitin ligase ITCH regulates life cycle of SARS-CoV-2 virus
    Article Snippet: .. HEK293 cells (ATCC, CRL-3216) at approximately 40–60% confluence were transfected with mixtures of plasmid DNA and cationic polymer linear polyethylenimine (593215, Polysciences). .. Vero and vT2 cells at approximately 40–60% confluence were transfected with mixtures of plasmid DNA and jetPRIME transfection reagent (101000046, Polyplus).

    Splicing Assay:

    Article Title: Investigating the molecular basis of the serological Rh D-negative phenotype in Indonesia: nature, frequency, and impact for diagnostics.
    Article Snippet: .. Minigene splicing assay (MSA) was carried out to investigate the effect of candidate SNVs on splicing as previously described.19–21 Briefly, short genomic regions consisting of RHD exon 3 (for c.486+5G>A) and 5 (for c.635-1G>A), as well as the flanking intronic regions, were PCR-amplified by using the gDNAs of interest as templates and were inserted into the minigene pSP3 vector by homologous recombination.20 After confirmation by sequencing, the wild-type and mutated recombinant vectors were transfected separately into HEK293 cells (ATCC Number: CRL-1573) and the fluorescent RT-PCR profiles expressed from the minigene constructs were analyzed by capillary electrophoresis as previously described.20 A ρSNV score was calculated to assess the direct effect of the variant on exon inclusion compared to the wild-type conditions (ρSNV = 1, no effect; ρSNV < 1, decrease in full-length exon inclusion; ρSNV = 0, no full-length exon inclusion).21 ..

    Polymerase Chain Reaction:

    Article Title: Investigating the molecular basis of the serological Rh D-negative phenotype in Indonesia: nature, frequency, and impact for diagnostics.
    Article Snippet: .. Minigene splicing assay (MSA) was carried out to investigate the effect of candidate SNVs on splicing as previously described.19–21 Briefly, short genomic regions consisting of RHD exon 3 (for c.486+5G>A) and 5 (for c.635-1G>A), as well as the flanking intronic regions, were PCR-amplified by using the gDNAs of interest as templates and were inserted into the minigene pSP3 vector by homologous recombination.20 After confirmation by sequencing, the wild-type and mutated recombinant vectors were transfected separately into HEK293 cells (ATCC Number: CRL-1573) and the fluorescent RT-PCR profiles expressed from the minigene constructs were analyzed by capillary electrophoresis as previously described.20 A ρSNV score was calculated to assess the direct effect of the variant on exon inclusion compared to the wild-type conditions (ρSNV = 1, no effect; ρSNV < 1, decrease in full-length exon inclusion; ρSNV = 0, no full-length exon inclusion).21 ..

    Sequencing:

    Article Title: Investigating the molecular basis of the serological Rh D-negative phenotype in Indonesia: nature, frequency, and impact for diagnostics.
    Article Snippet: .. Minigene splicing assay (MSA) was carried out to investigate the effect of candidate SNVs on splicing as previously described.19–21 Briefly, short genomic regions consisting of RHD exon 3 (for c.486+5G>A) and 5 (for c.635-1G>A), as well as the flanking intronic regions, were PCR-amplified by using the gDNAs of interest as templates and were inserted into the minigene pSP3 vector by homologous recombination.20 After confirmation by sequencing, the wild-type and mutated recombinant vectors were transfected separately into HEK293 cells (ATCC Number: CRL-1573) and the fluorescent RT-PCR profiles expressed from the minigene constructs were analyzed by capillary electrophoresis as previously described.20 A ρSNV score was calculated to assess the direct effect of the variant on exon inclusion compared to the wild-type conditions (ρSNV = 1, no effect; ρSNV < 1, decrease in full-length exon inclusion; ρSNV = 0, no full-length exon inclusion).21 ..

    Recombinant:

    Article Title: Investigating the molecular basis of the serological Rh D-negative phenotype in Indonesia: nature, frequency, and impact for diagnostics.
    Article Snippet: .. Minigene splicing assay (MSA) was carried out to investigate the effect of candidate SNVs on splicing as previously described.19–21 Briefly, short genomic regions consisting of RHD exon 3 (for c.486+5G>A) and 5 (for c.635-1G>A), as well as the flanking intronic regions, were PCR-amplified by using the gDNAs of interest as templates and were inserted into the minigene pSP3 vector by homologous recombination.20 After confirmation by sequencing, the wild-type and mutated recombinant vectors were transfected separately into HEK293 cells (ATCC Number: CRL-1573) and the fluorescent RT-PCR profiles expressed from the minigene constructs were analyzed by capillary electrophoresis as previously described.20 A ρSNV score was calculated to assess the direct effect of the variant on exon inclusion compared to the wild-type conditions (ρSNV = 1, no effect; ρSNV < 1, decrease in full-length exon inclusion; ρSNV = 0, no full-length exon inclusion).21 ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Investigating the molecular basis of the serological Rh D-negative phenotype in Indonesia: nature, frequency, and impact for diagnostics.
    Article Snippet: .. Minigene splicing assay (MSA) was carried out to investigate the effect of candidate SNVs on splicing as previously described.19–21 Briefly, short genomic regions consisting of RHD exon 3 (for c.486+5G>A) and 5 (for c.635-1G>A), as well as the flanking intronic regions, were PCR-amplified by using the gDNAs of interest as templates and were inserted into the minigene pSP3 vector by homologous recombination.20 After confirmation by sequencing, the wild-type and mutated recombinant vectors were transfected separately into HEK293 cells (ATCC Number: CRL-1573) and the fluorescent RT-PCR profiles expressed from the minigene constructs were analyzed by capillary electrophoresis as previously described.20 A ρSNV score was calculated to assess the direct effect of the variant on exon inclusion compared to the wild-type conditions (ρSNV = 1, no effect; ρSNV < 1, decrease in full-length exon inclusion; ρSNV = 0, no full-length exon inclusion).21 ..

    Construct:

    Article Title: Investigating the molecular basis of the serological Rh D-negative phenotype in Indonesia: nature, frequency, and impact for diagnostics.
    Article Snippet: .. Minigene splicing assay (MSA) was carried out to investigate the effect of candidate SNVs on splicing as previously described.19–21 Briefly, short genomic regions consisting of RHD exon 3 (for c.486+5G>A) and 5 (for c.635-1G>A), as well as the flanking intronic regions, were PCR-amplified by using the gDNAs of interest as templates and were inserted into the minigene pSP3 vector by homologous recombination.20 After confirmation by sequencing, the wild-type and mutated recombinant vectors were transfected separately into HEK293 cells (ATCC Number: CRL-1573) and the fluorescent RT-PCR profiles expressed from the minigene constructs were analyzed by capillary electrophoresis as previously described.20 A ρSNV score was calculated to assess the direct effect of the variant on exon inclusion compared to the wild-type conditions (ρSNV = 1, no effect; ρSNV < 1, decrease in full-length exon inclusion; ρSNV = 0, no full-length exon inclusion).21 ..

    Electrophoresis:

    Article Title: Investigating the molecular basis of the serological Rh D-negative phenotype in Indonesia: nature, frequency, and impact for diagnostics.
    Article Snippet: .. Minigene splicing assay (MSA) was carried out to investigate the effect of candidate SNVs on splicing as previously described.19–21 Briefly, short genomic regions consisting of RHD exon 3 (for c.486+5G>A) and 5 (for c.635-1G>A), as well as the flanking intronic regions, were PCR-amplified by using the gDNAs of interest as templates and were inserted into the minigene pSP3 vector by homologous recombination.20 After confirmation by sequencing, the wild-type and mutated recombinant vectors were transfected separately into HEK293 cells (ATCC Number: CRL-1573) and the fluorescent RT-PCR profiles expressed from the minigene constructs were analyzed by capillary electrophoresis as previously described.20 A ρSNV score was calculated to assess the direct effect of the variant on exon inclusion compared to the wild-type conditions (ρSNV = 1, no effect; ρSNV < 1, decrease in full-length exon inclusion; ρSNV = 0, no full-length exon inclusion).21 ..



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    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 <t>HEK293</t> 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 = <t>human</t> <t>embryonic</t> <t>kidney;</t> miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.
    Hek293 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hek293 t cells  (ATCC)
    99
    ATCC hek293 t cells
    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 <t>HEK293</t> 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 = <t>human</t> <t>embryonic</t> <t>kidney;</t> miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.
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    hek293 cells  (Servicebio Inc)
    86
    Servicebio Inc hek293 cells
    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 <t>HEK293</t> 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.
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    Image Search Results


    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.

    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

    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.

    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

    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.

    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