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: A method for expressing two proteins at a tunable ratio allows for studying the kinetics of creatine transport by MCT12

doi: 10.1016/j.isci.2026.117719

Figure Lengend Snippet: Adaptable expression ratios of the creatine transporters CRT1 and MCT12 in HEK293 SLC6A8 −/− cells (A) Scheme of the CRT1 3xHA -P2A-K n -MCT12 2xmyc construct. (B) Immunoblot analysis of CRT1 3xHA -P2A-K n -MCT12 2xmyc . (C) Ratio of MCT12 2xmyc and CRT1 3xHA expression as determined by immunoblotting ( n = 5). Data are represented as mean ± SEM. (D) Immunoblot analysis of CRT1 3xHA -P2A-K n -MCT12 2xmyc . (E) Ratio of MCT12 2xmyc and CRT1 3xHA expression as determined by immunoblotting ( n = 4). Data are represented as mean ± SEM. See also .

Article Snippet: Human: HEK293 Lenti-X , Takara , Cat # 632180.

Techniques: Expressing, Construct, Western Blot, Analysis

Journal: iScience

Article Title: A method for expressing two proteins at a tunable ratio allows for studying the kinetics of creatine transport by MCT12

doi: 10.1016/j.isci.2026.117719

Figure Lengend Snippet: Relative cell surface expression of the creatine transporters CRT1 and MCT12 in HEK293 SLC6A8 −/− cells (A) Scheme of the CRT1 eGFP -P2A-K 0 -MCT12 mCherry (a) and MCT12 mCherry -P2A-K 0 -CRT1 eGFP construct (b). (B) Ratio of mCherry and eGFP expression in eGFP-P2A-K 0 -mCherry (K 0 , as a control), CRT1 eGFP -P2A-K 0 -MCT12 mCherry , and MCT12 mCherry -P2A-K 0 -CRT1 eGFP constructs as determined by flow cytometry ( n = 6). Data are represented as mean ± SEM. (C) Live-cell confocal microscopy of HEK293 SLC6A8 −/− cells expressing CRT1 eGFP -P2A-K 0 -MCT12 mCherry and MCT12 mCherry -P2A-K 0 -CRT1 eGFP . The plasma membrane was stained with CellMask Deep Red. Scale bars, 20 μm. (D) Analysis of cell surface expression of CRT1 eGFP and MCT12 mCherry . n = 30 (CRT1 eGFP -P2A-K 0 -MCT12 mCherry ); n = 28 (MCT12 mCherry -P2A-K 0 -CRT1 eGFP ).

Article Snippet: Human: HEK293 Lenti-X , Takara , Cat # 632180.

Techniques: Expressing, Construct, Control, Flow Cytometry, Confocal, Microscopy, Clinical Proteomics, Membrane, Staining, Analysis

Journal: iScience

Article Title: A method for expressing two proteins at a tunable ratio allows for studying the kinetics of creatine transport by MCT12

doi: 10.1016/j.isci.2026.117719

Figure Lengend Snippet:

Article Snippet: Human: HEK293 Lenti-X , Takara , Cat # 632180.

Techniques: Cell-Signaling, Virus, Recombinant, Clinical Proteomics, Membrane, Staining, Cloning, Software

Journal: Blood Advances

Article Title: Distinct epitope engagement confers differential activity of linvoseltamab vs teclistamab across BCMA mutations ∗

doi: 10.1182/bloodadvances.2026020381

Figure Lengend Snippet: Binding of linvoseltamab and teclistamab to WT or mutated BCMA expressed by engineered HEK293/hCD20 cells. (A) Histogram plots from cell staining with 100nM anti-BCMA clone 19F2 or mIgG isotype control. BCMA is expressed on all engineered cells. (B) Histogram plots from staining with 100nM linvoseltamab, teclistamab, or hIgG isotype control. Linvoseltamab binding to BCMA R27P, P33S, and S30del was comparable to WT BCMA. Teclistamab binding to R27P, S30del, and P34del was reduced. (C) EC 50 values (nM) derived from nonlinear regression based on the cell staining experiment. hIgG, human immunoglobulin G; mIgG, mouse immunoglobulin G; NC, not calculated (EC 50 cannot be calculated from the range of concentrations tested); ND, not determined (no concentration-dependent binding was observed).

Article Snippet: Human embryonic kidney 293 (HEK293) cells (ATCC CRL-1573) stably expressing human CD20 (hCD20) (MS4A1; M1-P297 of NP_690605.1 ) were generated by transfection with a mammalian expression vector and selected in media (Dulbecco's Modified Eagle Medium, 10% fetal bovine serum, and 1% penicillin-streptomycin; Thermo Fisher Scientific) supplemented with 0.5 mg/mL of G418.

Techniques: Binding Assay, Staining, Control, Derivative Assay, Concentration Assay

Journal: Blood Advances

Article Title: Distinct epitope engagement confers differential activity of linvoseltamab vs teclistamab across BCMA mutations ∗

doi: 10.1182/bloodadvances.2026020381

Figure Lengend Snippet: Effects of BCMA mutations on linvoseltamab- and teclistamab-mediated Jurkat-NFAT cell activation. Jurkat-NFAT reporter cells cocultured with HEK293/hCD20 cells with WT or mutated BCMA; reporter activity was detected as luminescence. A nontargeting CD3 antibody was used as a control. RLU, relative light units.

Article Snippet: Human embryonic kidney 293 (HEK293) cells (ATCC CRL-1573) stably expressing human CD20 (hCD20) (MS4A1; M1-P297 of NP_690605.1 ) were generated by transfection with a mammalian expression vector and selected in media (Dulbecco's Modified Eagle Medium, 10% fetal bovine serum, and 1% penicillin-streptomycin; Thermo Fisher Scientific) supplemented with 0.5 mg/mL of G418.

Techniques: Activation Assay, Activity Assay, Control

Journal: Blood Advances

Article Title: Distinct epitope engagement confers differential activity of linvoseltamab vs teclistamab across BCMA mutations ∗

doi: 10.1182/bloodadvances.2026020381

Figure Lengend Snippet: Effect of BCMA mutations on T-cell–mediated cytotoxicity against HEK293h/CD20 cells treated with linvoseltamab and teclistamab. HEK293 or OPM-2 myeloma cells were incubated for 3 days with isolated human T cells, with E:T of 12.5 (HEK293) and 5 (OPM-2). Target cell survival was determined by flow cytometry. Percentage survival was normalized to untreated cells. The OPM-2 cell line was used as a comparator because this is an MM cell line that endogenously expresses BCMA. E:T, effector-to-target ratio.

Article Snippet: Human embryonic kidney 293 (HEK293) cells (ATCC CRL-1573) stably expressing human CD20 (hCD20) (MS4A1; M1-P297 of NP_690605.1 ) were generated by transfection with a mammalian expression vector and selected in media (Dulbecco's Modified Eagle Medium, 10% fetal bovine serum, and 1% penicillin-streptomycin; Thermo Fisher Scientific) supplemented with 0.5 mg/mL of G418.

Techniques: Incubation, Isolation, Flow Cytometry

Journal: Blood Advances

Article Title: Distinct epitope engagement confers differential activity of linvoseltamab vs teclistamab across BCMA mutations ∗

doi: 10.1182/bloodadvances.2026020381

Figure Lengend Snippet: Effect of BCMA-binding interface mutations on primary T-cell activation by linvoseltamab and teclistamab. HEK293 or OPM-2 myeloma cells were incubated for 3 days with isolated human T cells, with E:T of 12.5 (HEK293) and 5 (OPM-2). T-cell activation was determined by flow cytometry based on the percentage of CD8 + T cells expressing CD25. The OPM-2 cell line was used as a comparator because this is an MM cell line that endogenously expresses BCMA.

Article Snippet: Human embryonic kidney 293 (HEK293) cells (ATCC CRL-1573) stably expressing human CD20 (hCD20) (MS4A1; M1-P297 of NP_690605.1 ) were generated by transfection with a mammalian expression vector and selected in media (Dulbecco's Modified Eagle Medium, 10% fetal bovine serum, and 1% penicillin-streptomycin; Thermo Fisher Scientific) supplemented with 0.5 mg/mL of G418.

Techniques: Binding Assay, Activation Assay, Incubation, Isolation, Flow Cytometry, 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 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