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

Article Title: T cell activation by antibody-like immunoreceptors: increase in affinity of the single-chain fragment domain above threshold does not increase T cell activation against antigen-positive target cells but decreases selectivity.

doi: 10.4049/jimmunol.173.12.7647

Figure Lengend Snippet: FIGURE 4. ErbB2 expression of target cells. Cells of ErbB2-expressing lines (SK-OV-3, Colo201, MCF-7, Colo320, and HaCat) and primary cells (PBL and keratinocytes) for the control (each 1 106 cells) were incubated with an anti-ErbB2 mAb or an isotype-matched control mAb. Bound Abs were detected by an FITC-conjugated anti-mouse IgG Ab, and cells were analyzed by flow cytometry. The MFI of bound Abs was determined and is expressed as arbitrary units. Due to their immunoreactivity, the cells were characterized as high (MFI, 100), medium (MFI, 10), and low (MFI, 10) ErbB2-positive or ErbB2-negative cells, respectively.

Article Snippet: SK-OV-3 (ATCC HTB77; American Type Culture Collection, Manassas, VA), MCF-7 (ATCC HTB22) Colo201 (ATCC CCL 224), and Colo320 (ATCC CCL 220.1) were derived from various adenocarcinomas expressing different amounts of ErbB2.

Techniques: Expressing, Control, Incubation, Cytometry

Journal: bioRxiv

Article Title: Antagonizing the serotonin receptor HTR2B drives antigen-specific cytotoxic T-cell responses and controls colorectal cancer growth

doi: 10.1101/2025.02.26.640476

Figure Lengend Snippet: TCGA-COAD data were analyzed. (A) Kaplan-Meier survival analysis of HTR 2B expression (derived from TCGA data) in 220 colon adenocarcinomas patient’s cohort. “HTR 2B -high” versus “HTR 2B -low” data were segregated based on the cohort’s top or bottom 25% with intratumoral HTR 2B expression. Mantel-cox log rank test. (B) Representative images of HTR 2B and Ki67 in the control (lamina propria) and lamina propria of colon adenocarcinoma (COAD) tissue. The original magnification is 200X, and the inset images are 600X. (C) Representative images of serotonin (5-HT), neurofilament-A (NF-A) in the control (lamina propria) and lamina propria of colon adenocarcinoma (COAD) tissue IHC. Original magnification 200X. (D) RNA-seq datasets (n=30 samples) were segregated into “HTR 2B -high” and “HTR 2B -low” (highest fifteen and lowest fifteen intra-tumoral HTR 2B expression), and the differential expression of selected genes was analyzed. (E) Principal component analysis (PCA) of “HTR 2B -high” versus “HTR 2B -low” datasets (n=15 patients/group). (F) Venn diagram showing numbers of mutually exclusive or shared genes in each group. (G) A volcano plot shows differentially expressed genes (DEGs) of “HTR 2B -high” versus “HTR 2B -low” datasets (n=15 patients/groups) are shown. (H) Heatmap statistics differentially expressed selected gene sets. (I) Gene ontology of altered biological processes. (J) Gene set enrichment analysis (GSEA) of the curated gene sets that affect the regulatory and effector functions of the T cells. (K) Intra-tumoral immune cell signatures were analyzed by deconvolution using TIMER2.0 of HTR 2B -high and -low datasets. (L) A TIDE computational analysis of the association between the tumor-infiltrating CD8 T cells (CTL) level (expression of CD8A, CD8B, GZMA, GZMB, and PRF1. Overall, patient survival with the intratumoral HTR 2B gene expression level. For each patient cohort, tumor samples were divided into HTR 2B -high (samples with HTR 2B expression one standard deviation above the average, shown in the left survival plot) and HTR 2B -low (remaining samples shown in the right survival plot) groups.

Article Snippet: The human colon adenocarcinoma tissue microarray (Novus Biological, Catalog # NBP2-78088) was stained with the antibodies and detected using chromogenic reagent DAB.

Techniques: Expressing, Derivative Assay, Control, RNA Sequencing, Gene Expression, Standard Deviation

Journal: Journal of cellular and molecular medicine

Article Title: Aryl Hydrocarbon Receptor Alleviates Hepatic Fibrosis by Inducing Hepatic Stellate Cell Ferroptosis.

doi: 10.1111/jcmm.70278

Figure Lengend Snippet: FIGURE 1 | AHR directly regulates Mrp1 transcription in mHSCs. (A–C) The expression of Ahr, Mrp1 or Cyp1a1 was detected by QPCR. (D) There are two potential AHR exogenous response elements on the Mrp1 promoter sequence. (E) Promoter sequences containing XREL1 and XREL2 were cloned onto PGL3 plasmids, and mutant plasmids were constructed. (F) Double luciferase assay to detect luciferase activity. (G) EMSA detects AHR binding to specific elements XREL1 and XREL2. (H) CHIP detects AHR binding to specific elements XREL1 and XREL2. Data are expressed as means ± SD; *p < 0.05, **p < 0.01 and ***p < 0.001; Student's t-test or one-way ANOVA.

Article Snippet: ChIP assays were performed using the ChIP Assay kit (Beyotime, P2078). mHSCs were treated with YH439 (5 μM) for 24 h. Subsequently, mHSCs were sonicated and then immunoprecipitated with the antibody against AHR (1:100 dilution BOSTER Cat# A00225- 4, RRID: AB_3095576) with IgG (1:100 dilution Proteintech Cat# 30000- 0- AP RRID AB_2819035) as a negative control.

Techniques: Expressing, Sequencing, Clone Assay, Mutagenesis, Construct, Luciferase, Activity Assay, Binding Assay