a 431 nuclear extract  (Santa Cruz Biotechnology)

Journal: The European Respiratory Journal

Article Title: Lrig1 expression identifies airway basal cells with high proliferative capacity and restricts lung squamous cell carcinoma growth

doi: 10.1183/13993003.00816-2020

Figure Lengend Snippet: N-nitroso-tris-chloroethylurea (NTCU)-induced murine lung squamous cell carcinoma. a) NTCU administration protocol for the induction of murine lung squamous cell carcinoma. b) Representative images of NTCU-induced lung lesions stained with haematoxylin and eosin. Flat atypia with enlarged, flattened nuclei and increased nuclear-cytoplasmic ratio. Low-grade dysplasia with presence of multiple, ordered layers of epithelial cells and clear organisation from the basement membrane to the luminal surface. Note flattened nuclei adjacent to the lumen. High-grade dysplasia showing disordered layers of epithelial cells and multiple enlarged nuclei. Invasive squamous cell carcinoma lesion, beginning to fill the alveolar spaces. Keratin pearl, a characteristic feature of squamous cell carcinoma. c) Immunofluorescence staining showing expression of the lung squamous cell carcinoma markers P63 and KRT5, and the proliferation marker Ki67, in NTCU-induced lesions. d) Antibody staining demonstrates lack of immunoreactivity for the lung adenocarcinoma marker surfactant protein C within NTCU-induced lesions. Basement membrane (dashed lines) and airway lumen (AL) are indicated. Scale bars=50 μm.

Article Snippet: The dorsal fur of 6-week-old female mice was shaved and 75 μL of 0.013 M N-nitroso-tris-chloroethylurea (NTCU) (Santa Cruz sc-212265) diluted in acetone applied twice weekly for 12 weeks.

Techniques: Staining, Membrane, Immunofluorescence, Expressing, Marker

Journal: The European Respiratory Journal

Article Title: Lrig1 expression identifies airway basal cells with high proliferative capacity and restricts lung squamous cell carcinoma growth

doi: 10.1183/13993003.00816-2020

Figure Lengend Snippet: Effects of leucine-rich repeats and immunoglobulin-like domains 1 ( Lrig1 ) loss-of-function on N-nitroso-tris-chloroethylurea (NTCU)-induced lung carcinogenesis. a) Immunoblot for LRIG1 on cerebellar tissue lysates from wild-type, Lrig1 -heterozygous and Lrig1 -null mice. Note that protein levels change in a gene-dosage dependent manner. α-tubulin is shown as a loading control. b) Weights of NTCU-treated and control animals (mean± sem , Lrig1 +/+ n=11, Lrig1 +/– n=9, Lrig1 –/– n=7). Two-way ANOVA: time effect (p<0.0001) and time × treatment interaction (p<0.0001). Arrow indicates significant differences between all treatment groups and control arm. Tukey's multiple comparisons test, p<0.05. c) Transverse lung sections of NTCU-treated mice stained with haematoxylin and eosin and processed for KRT5 immunohistochemistry to identify pre-invasive airway lesions and invasive tumours. d) Proportion of lower airway epithelium displaying abnormal KRT5 expression in wild-type (+/+), Lrig1 -heterozygous (+/−) and Lrig1 -null (−/−) mice treated with NTCU (mean± sem , Kruskal–Wallis test, p=0.033, followed by Dunn's multiple comparisons test, p=0.049; n=7–11 mice per group). e) KRT5-expressing lesions were categorised as flat atypia, low-grade or high-grade. Distribution of pre-invasive lesions of different grades within lower airway (mean± sem ). Two-way ANOVA: genotype effect (p=0.009) and lesion grade effect (p<1×10 −6 ). The presence of flat atypia was statistically different among genotypes (Tukey's multiple comparisons test, p=7.7×10 −4 , p=1.7×10 −5 ). f) Representative images of NTCU-induced pre-invasive lesions stained for KRT5. Scale bars=50 μm. g) Number of invasive tumours per mouse. Kruskal–Wallis indicated no significant differences among groups (n=7–11 mice per genotype). h) Size of invasive tumours in mice of different Lrig1 genotypes (mean± sem ). Kruskal–Wallis test, p=0.0002, followed by Dunn's multiple comparisons test, p=0.005, p=0.0003; Lrig1 +/+ n=23, Lrig1 +/– n=142, Lrig1 –/– n=43 invasive lesions. IHC: immunohistochemistry.

Article Snippet: The dorsal fur of 6-week-old female mice was shaved and 75 μL of 0.013 M N-nitroso-tris-chloroethylurea (NTCU) (Santa Cruz sc-212265) diluted in acetone applied twice weekly for 12 weeks.

Techniques: Western Blot, Control, Staining, Immunohistochemistry, Expressing

Journal: The European Respiratory Journal

Article Title: Lrig1 expression identifies airway basal cells with high proliferative capacity and restricts lung squamous cell carcinoma growth

doi: 10.1183/13993003.00816-2020

Figure Lengend Snippet: Characterisation of N-nitroso-tris-chloroethylurea (NTCU)-induced lesions. a) Expression of P63 in KRT5-expressing squamous airway lesions of different grades in wild-type, leucine-rich repeats and immunoglobulin-like domains 1 ( Lrig1 )-heterozygous and Lrig1 -null mice (mean± sem ). b) Ki67 immunoreactivity in KRT5-expressing squamous airway lesions of increasing grades in Lrig1 +/+ , Lrig1 +/– and Lrig1 –/– mice (mean± sem ). For each lesion grade, the effect of Lrig1 genotype on the fraction of KRT5 + cells expressing each marker was assessed. Kruskal–Wallis test, followed by Dunn's multiple comparisons test, *: p<0.05. For both a) and b) n=11–31 lesions/grade/genotype; KRT5 + cells: Lrig1 +/+ n=6776, Lrig1 +/– n=10 206, Lrig1 –/– n=12 845.

Article Snippet: The dorsal fur of 6-week-old female mice was shaved and 75 μL of 0.013 M N-nitroso-tris-chloroethylurea (NTCU) (Santa Cruz sc-212265) diluted in acetone applied twice weekly for 12 weeks.

Techniques: Expressing, Marker

Journal: Cell

Article Title: A Patient-Derived Glioblastoma Organoid Model and Biobank Recapitulates Inter- and Intra-tumoral Heterogeneity

doi: 10.1016/j.cell.2019.11.036

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Goat polyclonal anti-Nestin (C-20) , Santa Cruz Biotechnology , Cat# sc-21247; RRID: AB_650014.

Techniques: Recombinant, Red Blood Cell Lysis, Modification, Saline, Activation Assay, Blocking Assay, Plasmid Preparation, Reverse Transcription, Sterility, Injection, Staining, Imaging, Enzyme-linked Immunosorbent Assay, Library Quantification, RNA Sequencing, Sequencing, Software, Adhesive, Cell Counting