Journal: Scientific Reports

Article Title: Novel NF-κB reporter mouse for the non-invasive monitoring of inflammatory diseases

doi: 10.1038/s41598-023-29689-4

Figure Lengend Snippet: Generation of NF-κB luciferase reporter mice. ( A ) Construct of the NF-κB luciferase reporter gene cassette integrated into the ROSA26 locus. pCAG, synthetic promoter consisting of CMV enhancer fused to the chicken beta-actin promoter; RE (responsive element), four repeats of NF-κB binding sequence; Stop, three repeats of SV40 polyA; TAp, minimal TA promoter (including the TATA box from HSV-tk promoter); WRPE, the woodchuck hepatitis virus posttranscriptional regulatory element; pgk-neo, neomycin-resistant gene driven by pgk promoter. ( B ) Bioluminescence and fluorescence images of C57BL/6 wild-type mice and NF-κB-Luc mice (NKL) in the PMA-induced ear inflammation model. PMA (2 μg) was applied to the right ears, and 6 h later, bioluminescence and fluorescence signals were detected 5 min after D-luciferin (100 µl, 15 mg/ml in PBS) i.p. injection. Radiance ranges of parameters are provided in the figure. ( C ) Mean group luminescence intensities (n = 3 per group). Results are presented as means ± SEMs. ( D ) Representative images of HE stained vehicle or PMA-treated ears (scale bar = 100 μm). Asterisk (*); thickened dermis with infiltrated immune cells.

Article Snippet: NF-κB-Luc:Lyz2 mice (n = 3) were treated with 3% (w/v) DSS (36–50 kDa) (MP Biomedicals, OH, USA) in drinking water for 5 days.

Techniques: Luciferase, Construct, Binding Assay, Sequencing, Fluorescence, Injection, Staining

Journal: Scientific Reports

Article Title: Novel NF-κB reporter mouse for the non-invasive monitoring of inflammatory diseases

doi: 10.1038/s41598-023-29689-4

Figure Lengend Snippet: LPS-induced systemic inflammation applied to NF-κB-Luc mice. ( A , B ) Bioluminescence images of PBS or LPS injected NF-κB-Luc mice ( A ) and total abdominal luminescence fluxes ( B ). The images were obtained at different times from each group (n = 3) using VISQUE InVivo ART100. Radiance ranges are provided in the figure. ( C ) Serum levels of IL-1β, IL-6, and TNFα. Results are presented as means ± SEMs (n = 3). * p < 0.05, ** p < 0.01, nd not detected. ( D ) Representative expressions of luciferase in liver, lung, and kidney as determined by western blotting. All western blot images were cropped from the original films presented in .

Article Snippet: NF-κB-Luc:Lyz2 mice (n = 3) were treated with 3% (w/v) DSS (36–50 kDa) (MP Biomedicals, OH, USA) in drinking water for 5 days.

Techniques: Injection, Luciferase, Western Blot

Journal: Scientific Reports

Article Title: Novel NF-κB reporter mouse for the non-invasive monitoring of inflammatory diseases

doi: 10.1038/s41598-023-29689-4

Figure Lengend Snippet: Generation of cell-type specific enhanced reporter mice and its ex vivo applications. ( A ) Comparison of bioluminescence and fluorescence among genotypes. Radiance ranges of parameters are provided in the figure. ( B ) Luciferase levels in liver and peritoneal macrophages isolated from NF-κB-Luc mice (NKL), NF-κB-Luc:Alb mice (NKLA), and NF-κB-Luc:Lyz2 mice (NKLL). LE; long exposure. ( C , D ) Luciferase levels in BMDMs isolated from NKL or NKLL. BMDMs were treated with LPS (100 ng/ml) for different times ( C ) or co-treated with LPS (100 ng/ml) and BAY 11–7085 (10 μM) for 6 h ( D ). Protein expressions were detected by western blotting. ( E , F ) Luminescence intensities of primary mouse hepatocytes (PMHs) isolated from NKL or NKLA. PMHs were treated with TNFα (5 or 20 ng/ml) ( E ), or co-treated with TNFα (20 ng/ml) and BAY 11–7085 (10 μM) for 24 h ( F ). Luminescence intensities were measured using microplate reader, and results are presented as means ± SEMs (n = 5). * p < 0.05, ** p < 0.01, *** p < 0.001. MP peritoneal macrophage, BMDM bone marrow-derived macrophage, PMH primary mouse hepatocyte. All western blot images were cropped from the original films presented in .

Article Snippet: NF-κB-Luc:Lyz2 mice (n = 3) were treated with 3% (w/v) DSS (36–50 kDa) (MP Biomedicals, OH, USA) in drinking water for 5 days.

Techniques: Ex Vivo, Fluorescence, Luciferase, Isolation, Western Blot, Derivative Assay

Journal: Scientific Reports

Article Title: Novel NF-κB reporter mouse for the non-invasive monitoring of inflammatory diseases

doi: 10.1038/s41598-023-29689-4

Figure Lengend Snippet: Non-invasive monitoring of inflammatory signals in the IBD model. NF-κB-Luc:Lyz2 (NKLL) mice were treated with 3% (w/v) dextran sodium sulfate (DSS) in drinking water for 5 days (n = 3) ( A ) Abdominal bioluminescence images and ( B ) luminescence intensities. Radiance ranges are provided in the figure. Luminescence intensity increased time-dependently after DSS treatment. ( C ) Body weight change during DSS treatment. ( D ) Correlation between DAI scores and luminescence intensities. The correlation coefficient ( r ) is shown in the figure. ( E ) Representative images of HE staining of colon tissues from mice before (control) and after 5 days of DSS treatment (DSS) (Scale bar = 100 μm). Histopathological features are indicated in the figure. * p < 0.05, ** p < 0.01.

Article Snippet: NF-κB-Luc:Lyz2 mice (n = 3) were treated with 3% (w/v) DSS (36–50 kDa) (MP Biomedicals, OH, USA) in drinking water for 5 days.

Techniques: Staining

Journal: Scientific Reports

Article Title: Novel NF-κB reporter mouse for the non-invasive monitoring of inflammatory diseases

doi: 10.1038/s41598-023-29689-4

Figure Lengend Snippet: Non-invasive monitoring of inflammatory signals in the NASH model. NF-κB-Luc (NKL) and NF-κB-Luc:Alb (NKLA) mice were fed normal chow diet for 2 weeks or choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) for 1 or 2 weeks. (n = 3 per group) ( A ) Bioluminescence images of Chow diet group, CDAHFD 1w group, and CDAHFD 2w group for both genotype. The images were obtained at the end of experiments, and the bioluminescence intensities were represented as a graph. The experimental schedule is described in detail in Materials and Methods. ( B ) Serum aspartate aminotransferase (AST) levels and ( C ) body weight changes of NKL and NKLA mice. ( D ) Representative HE images of livers obtained from each group of NKL and NKLA mice (scale bar = 100 μm), and ( E ) histopathological scores. cv; central vein, arrowhead; inflammatory foci. ( F ) Correlations between histopathological scores and luminescence intensities. Correlation coefficients ( r ) are shown in the graphs. ns not significant.

Article Snippet: NF-κB-Luc:Lyz2 mice (n = 3) were treated with 3% (w/v) DSS (36–50 kDa) (MP Biomedicals, OH, USA) in drinking water for 5 days.

Techniques: