Journal: Methods and Protocols

Article Title: Novel In Vivo Mouse Cryoablation Model to Explore Unique Therapeutic Approaches for Premalignant Columnar Lesions

doi: 10.3390/mps4010006

Figure Lengend Snippet: Preliminary results using the cryoablation procedure. ( A ) The experimental set-up of the validation of the cryoablation procedure is according to with minor adjustments. ( B ) Fluorescent image of untreated K5-GFP mice stomach. ( C ) Fluorescent image of vehicle-treated K5-GFP mice, 14 days post-cryoablation. ( D ) Fluorescent image of K5-GFP mice, 14 days post-cryoablation, treated with mNoggin (orally, 1× day). The delimitation of the original SCJ that separated the squamous and the columnar epithelium before the ablation is indicated by the dotted white line. i.p.: intraperitoneal injection; PO: Per os. , oral administration.

Article Snippet: Phosphate-buffered saline (PBS) (Thermo Fisher Scientific, Waltham, MA, USA; cat. no.: 10010023); Formalin solution, neutral buffered, 10% (Sigma-Aldrich, St. Louis, MO, USA; cat. no.: HT501128-4L); Ethanol (Sigma-Aldrich, St. Louis, MO, USA; cat. no.: 51976-500ML-F); Xylene (Sigma-Aldrich, St. Louis, MO, USA; cat. no.: 108298); Hematoxylin solution according to Mayer (Sigma-Aldrich, St. Louis, MO, USA; cat. no.: 51275); Eosin Y solution 1% (Sigma-Aldrich, St. Louis, MO, USA; cat. no.: 1170811000); DPX Mountant (Sigma-Aldrich, St. Louis, MO, USA; cat. no.: 06522-100ML); Tissue-Plus TM O.C.T. compound (Fisher Scientific, Waltham, MA, USA; cat. no.: 23-730-571); DAPI (4’,6-diamidino-2-phenylindole, dihydrochloride) (Thermo Fisher Scientific, Waltham, MA, USA; cat. no.: 62247); Recombinant mouse noggin protein (R&D Systems, McKinley, MN, USA; cat. no.: 1967-NG); Tamoxifen (Sigma-Aldrich, St. Louis, MO, USA; cat. no.: T5648).

Techniques: Biomarker Discovery, Injection

Journal: Scientific Reports

Article Title: Human TNF-Luc reporter mouse: A new model to quantify inflammatory responses

doi: 10.1038/s41598-018-36969-x

Figure Lengend Snippet: Effect of anti-inflammatory IL-10 on luciferase activity in hTNF.LucBAC BMDMs. BMDMs were either unstimulated (dotted line) or stimulated with 10 ng/mL LPS, in the absence (black line) or in the presence of increasing levels of recombinant murine IL-10, and luciferase activity was monitored over time; IL-10 concentrations 0.01 (orange), 0.1 (blue), 1 (green) and 10 ng/mL (red) ( a ). The ED50 of IL-10 as determined by the dose-dependent inhibition of luciferase activity in stimulated hTNF.LucBAC BMDMs was 0.6 ng/mL ( b ). Soluble TNF detected in the medium 24 h after LPS stimulation in the absence or presence of IL-10 ( c ). Correlation of IL-10 blockade of LPS-induced luciferase activity represented as area under the curve (AUC) with secreted mouse TNF ( d ). IKK-2 inhibitor BI605906 (BI) was used as a positive control for inhibition of luciferase activity and soluble TNF release. BMDMs left unstimulated (dotted line) or treated with 10 ng/mL of LPS, in the absence (black line) or presence of BI inhibitor at 0.01 μM (orange), 0.1 μM (blue), 1 μM (green) and 10 μM (red) ( e ). Luciferase activity (AUC) in the absence or presence of BI in LPS-induced cells (f ). Data are representative of three independent experiments, with triplicate cultures per experiment (N = 3, n = 3) and bars represent standard error of the mean.

Article Snippet: The crypt pellet was resuspended in ice-cold Matrigel (Scientific Laboratory Supplies, UK) (500 crypts/50 μL Matrigel) containing 50 mg/mL recombinant mouse epidermal growth factor (EGF), 100 ng/mL recombinant mouse Noggin CF and 500 ng/mL recombinant mouse R-spondin 1; all obtained from R&D Systems UK.

Techniques: Luciferase, Activity Assay, Recombinant, Inhibition, Positive Control