firefly luciferase assay kit  (Biotium)

Journal: Bone Research

Article Title: Coactivator-independent vitamin D receptor signaling causes severe rickets in mice, that is not prevented by a diet high in calcium, phosphate, and lactose

doi: 10.1038/s41413-024-00343-7

Figure Lengend Snippet: Increased corepressor but no coactivator interaction with VDR ΔAF2 . a Log ten-fold change (LFC) difference in coregulator interactions between 1,25(OH) 2 D 3 - vs DMSO-treated VDR +/+ and VDR ΔAF2 proteins, measured by NAPing and ordered by hierarchical clustering (Euclidean distance, average linkage). b Absolute arbitrary unit of fluorescence as a measure of coregulator binding to the indicated (mutant) VDR, depicted for the indicated (black arrowheads) peptides of NCOA1, NCOR1, and NCOR2. Student’s t-test 1,25(OH) 2 D 3 vs DMSO. Results are expressed as mean ± SD (4 technical replicates). c Luciferase transactivation assay to determine the 1,25(OH) 2 D 3 -induced transactivation capacity of the indicated (mutant) receptors, analyzed by two-way ANOVA followed by Dunnett multiple comparisons test of (mutant) receptor vs baseline. Results are expressed as mean ± SEM, (2 independent experiments with 3 technical replicates each)

Article Snippet: After 24 h, luciferase activity was measured with a Firefly luciferase kit (Biotium, VWR, Avantor) and normalized to β-galactosidase activity, measured with the Galacto-Light Plus System (Applied Biosystems).

Techniques: Fluorescence, Binding Assay, Mutagenesis, Luciferase, Transactivation Assay

Journal: Advanced healthcare materials

Article Title: STING protein-based in situ vaccine synergizes CD4 + T, CD8 + T and NK cells for tumor eradication

doi: 10.1002/adhm.202300688

Figure Lengend Snippet: (A) Protein homology-modeling structural prediction of dimerized anti-PDL1-STINGΔTM protein complexed with a cGAMP molecule with SWISS modeling. (B) HEK293T interferon-luciferase reporter cells (N = 3) treated with different ICB-ΔTM-cGAMP complexes, along with S365AΔTM mutant controls. Luciferase activity was determined 24 hours post treatment. (C) Ex vivo assay of OT1 CD8+ T cells’ cytotoxicity against SIINFEKL-MC38 cancer cells, ICB-ΔTM-cGAMP complexes along with equal molar amount of anti-PDL1 antibody were added during the co-culture of CD8+ T cells and cancer cells, the cytotoxicity was measured with flow cytometry with representative plot shown in (D). (E) Representative IVIS images of biodistribution (F) of Cy5-labeled ICB-ΔTM-cGAMP complexes 24 hours post intratumoral injections (tdLN for tumor draining lymph node and cLN for contralateral lymph node). Data were analyzed with one-way ANOVA.

Article Snippet: 24 hours post treatment, cells were lysed for firefly luciferase assay (Biotium, Cat#: 30075–2) to quantify the interferon-luciferase activity.

Techniques: Luciferase, Mutagenesis, Activity Assay, Ex Vivo, Co-Culture Assay, Flow Cytometry, Labeling