Journal: International Journal of Molecular Medicine

Article Title: Recombinant ADAMTS-1 promotes muscle regeneration accompanied by downregulation of Notch signaling

doi: 10.3892/ijmm.2025.5718

Figure Lengend Snippet: Transcriptomic analysis and GO enrichment of muscle cell differentiation following rADAMTS-1 treatment. C2C12 cells were treated with 10 ng/ml rADAMTS-1 and differentiated for 3 days before RNA-seq analysis. (A) Volcano plot showing DEGs. Blue and red dots indicate significantly upregulated and downregulated DEGs, respectively. (B) Validation of selected DEGs from RNA-sequencing by quantitative PCR. Results are presented as mean ± SD. Statistical significance was determined using one-way analysis of variance with Tukey's post hoc test and is indicated as follows: * P<0.05 and ** P<0.01 vs. the control group. GO enrichment analysis of the top 20 terms associated with (C) Biological Process, (D) Cellular Component and (E) Molecular Function, based on the identified DEGs. GO, Gene Ontology; ADAMTS-1, a disintegrin and metalloproteinase with thrombospondin motifs 1; rADAMTS-1, recombinant ADAMTS-1; RNA-seq, RNA sequencing; DEG, differentially expressed genes.

Article Snippet: Mouse myoblast C2C12 cells (cat. no. CRL-1772; ATCC) were obtained from ATCC.

Techniques: Cell Differentiation, RNA Sequencing, Biomarker Discovery, Real-time Polymerase Chain Reaction, Control, Recombinant

Journal: Scientific Reports

Article Title: Influenza A virus derived NS1 enhances translation of HPLC purified mRNA and interferon adjuvanted mRNA vaccination

doi: 10.1038/s41598-026-35611-5

Figure Lengend Snippet: ( A ) C2C12 cells (n=5) were seeded in 24 well plates and co-transfected with Ψ modified mRNAs encoding luciferase (L) , GFP (G) and PR8S (P) at mass ratios L G P = 1 : (1−ε): ε (where ε is 0, 0.5 or 1) . 4 h after mRNA complexes were added, cells were treated with cytokines supplemented media at non-toxic concentrations IFN-β (50ng/mL), IL-1β (1ng/mL) or IL-6 (1ng/mL) for 18 h followed by luciferase assay. Cell viability of all groups were similar (Supplementary Figure S5). Results were normalized by luciferase expression of non-treated cells and reported as relative luciferase expression representing extent of translation inhibition by cytokines. Results were presented as the mean SEM. ( B ) C2C12 cells (n=5) were co-transfected with Ψ modified mRNAs encoding luciferase (L), IFN-α (I), GFP (G) and PR8S (P) at mass ratios L I G P = 2 : 1 : (1−ε): ε (where ε is 0, 0.5, 0.75 or 1). Control cells for each mass ratio were similarly transfected with G replacing I i.e. L G G P = 2 : 1: (1−ε) : ε. Luciferase expression was measured 18h later. Cell viability of all groups were similar (Supplementary Figure S5). Results were presented as the mean SEM. ( C ) C57Bl/6 Mice (n=8) were intramuscularly injected with 8 µg Luc mRNA, together with 8 µg GFP mRNA (Luc+GFP), 4 µg GFP mRNA + 4 µg IFN-α mRNA (Luc+GFP+IFN), 4 µg GFP mRNA + 4 µg PR8S mRNA (Luc+PR8S+GFP), or 4 µg PR8S mRNA + 4 µg IFN-α mRNA (Luc+PR8S+IFN). Non-treated (NT) group was injected with buffer without mRNA. Bioluminescence was assayed by IVIS 5 h after injection and once daily thereafter. Results were presented as the mean SEM. ( D ) C57Bl/6 Mice (n=4) were intramuscularly injected twice (7 days apart) with 8 µg OVA mRNA, together with either 8 µg GFP mRNA (OVA+GFP), 4 µg GFP mRNA + 4 µg IFN-α mRNA (OVA+GFP+IFN), 4 µg PR8S mRNA + 4 µg GFP mRNA (OVA+PR8S+GFP), 4 µg PR8S mRNA + 4 µg IFN-α mRNA (OVA+PR8S+IFN). Non-treated (NT) group was injected with buffer without mRNA. 7 days after the second injection, blood was collected by cardiac puncture and anti-OVA IgG titers were quantified by ELISA. Results were presented as the mean SEM with individual values shown.

Article Snippet: NIH-3T3 mouse fibroblasts, C2C12 mouse myoblast cell line and HepG2 liver cancer cells were purchased from the American Type Culture Collection (ATCC); NIH-3T3 cells were cultured in Dulbecco’s Modified Eagle Medium (Hyclone).

Techniques: Transfection, Modification, Luciferase, Expressing, Inhibition, Control, Injection, Enzyme-linked Immunosorbent Assay