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

Journal: RSC Chemical Biology

Article Title: Signatures of native-like glycosylation in RNA replicon-derived HIV-1 immunogens

doi: 10.1039/d5cb00165j

Figure Lengend Snippet: Differential glycan composition on Env expressed in different cell lines. Glycan composition at reporter glycosylation sites predominantly displaying: oligomannose-type glycans (N332), a mixture of complex- and oligomannose-type glycans (N355), and complex-type glycans (N88). Four different production systems were used as described in the key, including expression in three different cell lines, HEK293F, C2C12 and DC2.4 cells. All the glycan composition observed in the site-specific analysis are shown on the left, simplified in distinct categories represented as, core, oligomannose-type, and complex-type glycans. Some compositions annotated as complex-type glycans can exhibit isomers formally constituting hybrid-type glycans. Represented glycan compositions (See Methods for full classification) are colored according to the scale provided in the right for each category of glycan composition. The representative glycan composition data at all the sites is the average of three or more biological replicates.

Article Snippet: The mouse muscle cell line (C2C12) was cultured in Dulbecco's modified Eagles medium supplemented with 10% fetal bovine serum (FBS) as recommended in manufacturer's protocol (American type culture collection, Catalogue no. CRL-1772).

Techniques: Glycoproteomics, Expressing