Journal: Immunity

Article Title: T cell-derived interleukin-22 drives the expression of CD155 by cancer cells to suppress NK cell function and promote metastasis

doi: 10.1016/j.immuni.2022.12.010

Figure Lengend Snippet: CD226 expression is higher on NK cells in Il22 −/− mice (A) Intravenous model of 4T1-luciferase + (4T1-Luc) metastasis. (B) Representative IVIS images and average radiance from one experiment of two (n = 5). (C) Ex vivo imaging of the lung by IVIS. Linear regression analysis of clonogenic colonies vs. average radiance. (D) Representative dot plots and gating strategy of IFNγ-producing cells in the lungs. Numbers represent the frequency of the parent gate. Frequency of IFNγ + cells in the lung and composition by cell type including CD8 + and CD4 + T cells, NKT cells, and NK cells. (E) Representative dot plots and frequency of IFNγ + NK cells (n = 9 and 10). Data from two independent experiments were pooled. (F) Representative fluorescent microscopy images of lungs from 4T1-injected mice. NKp46 FITC signal is depicted in red, CD8 AF555 in yellow, CD155 PE in green, and Hoechst DNA staining in blue. Intralesional CD155 PE MFI per lesion in WT and Il22 −/− mice. Chip cytometry spatial distribution of NKp46 + (red), CD8 + (green), and CD155 + (gray) cells in the microscopy samples. Frequency of CD8 and NKp46 cells per lesion from the previous graph. Data presented as means ± SEM; p values <0.05 were considered significant by paired t test of one chip of 2 for each condition (n = 3 and 4). (G) Gating strategy and representative plots of CD226 staining on CD8 + T, CD4 + T, NKT, and NK cells. Isotype control is in black. Frequency of CD226 + cells in the lungs (n = 6). Data of one experiment of two. Data presented as means ± SEM; p values <0.05 were considered significant by the Mann-Whitney U test or multiple unpaired t tests with Holm-Sidak correction. See also Figure S6 .

Article Snippet: The following day, FITC conjugated donkey anti-goat antibody (Jackson ImmunoResearch) was applied at a concentration of 10 μg/mL for 2 h at room temperature.

Techniques: Expressing, Luciferase, Ex Vivo, Imaging, Microscopy, Injection, Staining, Chip Cytometry, Control, MANN-WHITNEY

Journal: Immunity

Article Title: T cell-derived interleukin-22 drives the expression of CD155 by cancer cells to suppress NK cell function and promote metastasis

doi: 10.1016/j.immuni.2022.12.010

Figure Lengend Snippet:

Article Snippet: The following day, FITC conjugated donkey anti-goat antibody (Jackson ImmunoResearch) was applied at a concentration of 10 μg/mL for 2 h at room temperature.

Techniques: Purification, Control, Virus, Recombinant, Red Blood Cell Lysis, Transfection, Flow Cytometry, DNA Extraction, Gel Extraction, Plasmid Preparation, cDNA Synthesis, Cell Differentiation, Staining, Cell Isolation, Conjugation Assay, Sequencing, CRISPR, Retroviral, Software

Journal: Immunity

Article Title: T cell-derived interleukin-22 drives the expression of CD155 by cancer cells to suppress NK cell function and promote metastasis

doi: 10.1016/j.immuni.2022.12.010

Figure Lengend Snippet:

Article Snippet: InVivoMAb rat IgG2a (RG7/1.30) , BioXcell , Cat# BE0251; RRID: AB_2687732.

Techniques: Purification, Control, Virus, Recombinant, Red Blood Cell Lysis, Transfection, Flow Cytometry, DNA Extraction, Gel Extraction, Plasmid Preparation, cDNA Synthesis, Cell Differentiation, Staining, Cell Isolation, Conjugation Assay, Sequencing, CRISPR, Retroviral, Software

Journal: Carcinogenesis

Article Title: Kras mutations and PU.1 promoter methylation are new pathways in murine radiation-induced AML

doi: 10.1093/carcin/bgz175

Figure Lengend Snippet: Analysis of Flt3 -ITD in murine rAMLs. ( A ) Agarose gel electrophoresis of Flt3 -ITD PCR of a panel of murine rAMLs on a 2% gel. A normal amplicon is represented by a single band of 333 bp whereas Flt3 -ITD has an additional larger band. The gel is loaded as indicated in the image. A cropped version of the gel is displayed. `CBA' and `C57' refer to normal spleen DNA from CBA/H and C57BL6, respectively; `Brain' refers to brain tissue from the animal in which AML A developed and represents a normal tissue control; and AMLs E–G refer to independent AML samples. Flt3 -ITDS in AMLs C and D have been previously analysed in Finnon et al. . The presence or absence of an Flt3 -ITD and chromosome 2 deletion are stated below the gel for each sample ( Flt3 -ITD: + presence and − absence; del2: + presence and − absence. ( B ) CBA sequence of exon 14 on chromosome 5 showing Flt3 -ITDs in AML A at nucleotide 2017 (24 bp) and in AML B at nucleotide 2035 (33 bp). ( C ) Predicted CBA Flt3 protein sequence with ITDs of AML A (aa.598) and AML B (aa.604).

Article Snippet: For the PCR reaction, 1 μl of extracted DNA (25 ng DNA/μl) was placed in 9 μl PCR master mix [1 μl ×10 PCR buffer, 2 μl ×5 Q-solution, 1.6 μl 1.25 mM dNTP, 3.15 μl dH 2 O, 0.5 μl 10 μM primer ‘Forward’, 0.5 μl 10 μM primer ‘Reverse’, 0.25 μl Taq DNA Polymerase (Qiagen, Manchester, UK).

Techniques: Agarose Gel Electrophoresis, Amplification, Control, Sequencing