hekblue htlr5 reporter cell  (InvivoGen)

Journal: NPJ Vaccines

Article Title: Antigen cross-presentation potentiating cancer vaccine adjuvant for T cell expansion and synergy with anti-PD-1

doi: 10.1038/s41541-026-01376-1

Figure Lengend Snippet: A The vector map of recombinant protein constructs: FMS-like tyrosine kinase-3 ligand extracellular domain (Flt3L/F), Vibrio vulnificus flagellin B (FlaB/B). The fusion protein was generated by combining F and B with B position at the C-terminal (FB) or the N-terminal (BF). B Characterization of the recombinant proteins by sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) and subsequent Western blot analysis using mouse anti-FlaB serum or rabbit anti-Flt3L antibody. C Determination of TLR5-dependent NF-κB stimulating activity of B, FB, and BF in various protein concentrations using HEK-Blue™ hTLR5 cells ( n = 3 biological replicates). D Total cell number and the number of cell cluster formations of BMDCs after 9 days of cultures supplemented with B, F, and FB. Floating cells were collected and counted using a Vi-Cell Blue machine ( n = 7 biological replicates), while cell clusters were visualized with an EMOS5000 machine ( n = 5 biological replicates). E Representative flow histogram of CD103 expression on CD11c + MHCII hi population and quantification data of CD103 + DCs generation by recombination protein in a dose-dependent manner using BMDCs ( n = 3 biological replicates). F Representative levels of CFSE in CD8 + T cells from cultures with the indicated DCs, the individual peaks of CFSE dilution gated, and quantitative proliferation index of CD8 + T cell. For all sections: **** P < 0.0001; *** P < 0.001; ** P < 0.01; * P < 0.05.

Article Snippet: To assess the TLR5-stimulating activities of the recombinant fusion proteins, we evaluated the TLR5-dependent NF-κB-stimulating activity of the recombinant protein using HEK-BlueTM hTLR5 cells (hκb-help-5, InvivoGen, USA) and HEK-BlueTM Detection (hb-det2, InvivoGen, USA) assay systems following the manufacturer’s instructions.

Techniques: Plasmid Preparation, Recombinant, Construct, Generated, SDS Page, Western Blot, Activity Assay, Expressing

Journal: Gut Microbes

Article Title: The response regulator FpsR controls the flagella–pili transition and mucosal colonization in Ligilactobacillus ruminis

doi: 10.1080/19490976.2025.2596807

Figure Lengend Snippet: Flagellin expression and proinflammatory activity of L. ruminis Mot strain. a, Profiling of surface-associated proteins extracted from L. ruminis . Mk, protein size marker; H-FliC, purified His-tagged FliC1/2. b, Western blot analysis using anti-FliC1/2 antibodies. c , Purified flagellins from various bacterial species: Lag, L. agilis ; Lru, L. ruminis Mot strain; ST, Salmonella Typhimurium ; LM, Listeria monocytogenes . d–f, Reporter gene assays using HEK-Blue hTLR5 cells stimulated with whole cells (d), intact purified flagella (e), or monomerized purified flagellin (f). The secreted alkaline phosphatase (SEAP) was measured. g–i, IL−8 production by Caco−2 cells stimulated with whole cells (g), intact purified flagella (h), or monomerized purified flagellin (i). IL−8 levels in the culture supernatants were quantified. n = 3; data are shown as mean ± standard error (SE). d, g, Significant difference between WT and Mot strains is indicated using an asterisk (* P < 0.0001; Two-way ANOVA with Bonferroni multiple comparisons test.).

Article Snippet: Proinflammatory responses were assessed based on previously described methods, including quantification of IL−8 production by Caco−2 cells and a reporter gene assay using HEK-Blue hTLR5 cells (InvivoGen).

Techniques: Expressing, Activity Assay, Marker, Purification, Western Blot