Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Toxicity induced by a bispecific T cell redirecting protein is mediated by both T cells and myeloid cells in immunocompetent mice 2

doi: 10.4049/jimmunol.1901401

Figure Lengend Snippet: (A) Diagram showing construction of treatment (mNKG2D-2C11) and control (TZ47-2C11) bsTCEs. (B-D) MC38 (1x106) cells were injected s.c. into WT B6 mice. Treatment was initiated when tumors reached approximately 40mm2. (B) Mice received a total of four 10μg i.v injections of bsTCE every other day. (Left) Health scoring was blinded and evaluated 3h and 24h after each treatment. (Right) Weight, normalized to day 0, at the indicated time points. Data pooled from 7 experiments (n=30) (C) Mice received a total of four i.v. injections of either 2, 5 or 10μg bsTCE every other day. Health scoring was blinded and evaluated every hour for the first 8h and then 24h after each treatment. Data pooled from 2 experiments (n=8). (D) Tumor or non-tumor bearing WT B6 mice received a total of four 10μg i.v injections of bsTCE every other day. Health scoring was blinded and evaluated 3h and 24h after each treatment. Data are pooled from two experiment (n=8). (E) Representative images of Rae1 positive staining in the liver of B6 mice or contiguous tissue sections blocked with recombinant Rae1. Slides counterstained with methylene blue. Health scores in B and D are shown +/− SD, weight and C are shown +/− SEM. Statistical significance determined by repeated measures two-way ANOVA with Bonferroni’s multiple comparisons test (B, C) or with Tukey multiple comparisons test (D). * p<0.05, *** p<0.001, NS = not statistically significant.

Article Snippet: Contiguous sections from each sample were incubated with primary antibodies pre-incubated for 4 hours with recombinant mouse Rae1 Fc chimeric protein (R&D Systems, 1998-RA) at a 1:10 molar ratio as specificity controls.

Techniques: Control, Injection, Staining, Recombinant

Journal: Nature

Article Title: Epigenetic Silencing by SETDB1 Suppresses Tumor Intrinsic Immunogenicity

doi: 10.1038/s41586-021-03520-4

Figure Lengend Snippet: (a) Proportion of chromatin accessible sites (ATAC-seq) gained in Setdb1 KO LLC or B16 cells that are located within (red) or outside (grey) SETDB1 domains. (b) Proportion of ATAC-seq sites gained in Setdb1 KO LLC or B16 cells that coincide with promoters (light grey), distal TEs (red), or other promoter-distal sites (dark grey). Statistics by permutation testing. (c) Proportion of gained ATAC-seq sites at distal TEs in Setdb1 KO B16 cells that also gain H3K27 acetylation and resemble active enhancers. (d) Coordinate gain of chromatin accessibility and H3K27 acetylation at an example TE-site in Setdb1 KO B16 cells. (e) Activation of genes near (<50kb) gained ATAC-seq sites at distal TEs in Setdb1 KO LLC or B16 cells compared to control genes. Statistics by permutation testing. (f-h) Flow cytometry in control and Setdb1 KO cells showing (f) gating strategy, (g) cell-surface expression (y-axis, median fluorescence intensity (MFI)) for ULBP1 and RAET1 ligands in LLC (left), and MHC-I expression in LLC and B16 (right) +/− induction with IFNγ (10ng/mL, 24hr). Data are mean +/− s.e.m. and reflect 2 independent experiments with 4 biological replicates. Statistics by two-sided Student’s t-test. *P < 0.05; **P < 0.01.

Article Snippet: Cells were stained with hybridoma supernatant against MuLV envelope proteins (ATCC, HB-10392) followed by secondary staining with fluorescent antibodies, or with directly conjugated fluorescent antibodies against ULBP1 (FAB2588P, R&D systems), pan-RAET1 (FAB17582P, R&D systems), or MHC Class I (28–8-6, BioLegend) at 1:100 dilution 50 .

Techniques: Activation Assay, Control, Flow Cytometry, Expressing, Fluorescence

Journal: Cell

Article Title: Natural Killer Cells Degenerate Intact Sensory Afferents following Nerve Injury

doi: 10.1016/j.cell.2018.12.022

Figure Lengend Snippet:

Article Snippet: RRID: AB_10011569 ), goat anti-mouse pan-RAE1 (1:40; R&D systems, cat no. AF1136.

Techniques: Control, Purification, Plasmid Preparation, Recombinant, Cell Isolation, LDH Cytotoxicity Assay, Enzyme-linked Immunosorbent Assay, RNAscope, HD Assay, Sequencing, Negative Control, Software