Journal: PeerJ

Article Title: BNC1 inhibits the development and progression of gastric cancer by regulating the CCL20/JAK-STAT axis

doi: 10.7717/peerj.19477

Figure Lengend Snippet: (A) Heatmap of gene expression changes after overexpression of BNC1. (B) Venn diagram showing the screening of target genes of BNC1. Genes showing decreased expression after overexpression of BNC1 and highly expressed genes in gastric cancer from the TCGA database. (C) Downstream target genes of BNC1 were screened by transcriptome sequencing in AGS stable cell lines overexpressing BNC1 and control cells (log 2 fold change [FC] > 1.5 or < −1.5). (D–H) mRNA levels of PTPRC, SERPINB9, RAC2, BAKN1, and CCL20 in MKN-28, AGS and GES-1 cell lines. (I) KEGG analysis of genes showing decreased expression after overexpression of BNC1. (J) mRNA levels of CCL20 in MKN-28 and AGS stable cell lines overexpressing BNC1 and in control cells. (K) ELISA shows the protein levels of CCL20 in MKN-28 and AGS cell lines after overexpression of BNC1. (L) AlphaFold3 prediction of BNC1 binding to the CCL20 promoter region. (M) Dual luciferase results for BNC1 and CCL20.

Article Snippet: The stable BNC1-overexpressing MKN-28 and AGS cell lines were used to perform the ELISA analysis using the Human MIP-3α/CCL20 (C-C motif chemokine ligand 20) QuickTest ELISA Kit (QT-EH0231; Wuhan Fine Biotech Co., Ltd).

Techniques: Gene Expression, Over Expression, Expressing, Sequencing, Stable Transfection, Control, Enzyme-linked Immunosorbent Assay, Binding Assay, Luciferase

Journal: Nature Communications

Article Title: Brucella abortus impairs T lymphocyte responsiveness by mobilizing IL-1RA-secreting omental neutrophils

doi: 10.1038/s41467-024-55799-2

Figure Lengend Snippet: a At 8 or 30 days post-infection (p.i.) of C57BL/6 mice i.p. infected with B. abortus (Ba) (red square) or mock-injected with PBS (black circle), omentum and peritoneal fluid were harvested, and individualized cells analyzed by flow cytometry. Normalized absolute numbers of omental and peritoneal F4/80 hi LPM, F4/80 int MHCII hi SPM, Ly6C hi monocytes, or Ly6G + neutrophils at the indicated time points are shown. Each symbol represents one animal. (PBS, 2-3 mice; Ba, 3-4 mice), n = 3. Mean from pooled data ± SD. Significant differences are shown (Mann-Whitney test, two-sided). Absence of p -value, non-significant. b Confocal microscopy images of wholemount staining of omenta harvested at day 8 or 30 p.i. from i.p. infected C57BL/6 mice or mock-PBS treated. F4/80 (green, macrophages), Ly6G (magenta, neutrophils), Ly6C (violet, monocytes and endothelial cells of blood vessels), B220 (red, B cells), and CD3 (cyan, T cells). B and/or T cell localization defines the milky spot (MS). Individual stainings of cells present in the selected square are on the right. 3 mice, n = 2. Scale bar, 20 µm. c Median expression of selected differentially expressed genes from published RNA seq transcriptomic profiling obtained from whole blood samples from healthy donor controls or primary brucellosis patients in acute, acute relapse, or chronic phase of infection , . X -axis: Healthy donor controls n = 32 (Gray); Brucellosis patients: Acute n = 56 (Yellow), Acute with relapse n = 6 but for CXCL1 n = 4 (Orange), Chronic n = 11 (Blue). Y-axis: log2 residual gene expression counts. Each symbol represents one patient or healthy donor. Mean ± SD. Significant differences are shown (Multiple comparison Kruskal Wallis test, followed by post-hoc Dunn’s test). Absence of p -value, non-significant. Source data are provided as a Source Data file.

Article Snippet: The dosage of murine CXCL1 (Cusabio, CSBE17286m) or IL-1RA (Thermofisher, EMIL1RN) was done by ELISA following the manufacturer’s instructions.

Techniques: Infection, Injection, Flow Cytometry, MANN-WHITNEY, Confocal Microscopy, Staining, Expressing, RNA Sequencing, Gene Expression, Comparison

Journal: Nature Communications

Article Title: Brucella abortus impairs T lymphocyte responsiveness by mobilizing IL-1RA-secreting omental neutrophils

doi: 10.1038/s41467-024-55799-2

Figure Lengend Snippet: a Serum, peritoneal fluid, and omentum were harvested from C57BL/6 mice at 48 h (filled circles) or 72 h (empty circles) after i.p. injection with Ba-wt LPS (red circles) or PBS (black circles). CXCL1 was measured by ELISA. Mean of pooled data from normalized absolute number concentrations ± SD. 2 mice per group, n = 2. b Confocal microscopy images of whole mount staining of omenta harvested at 4 h and 72 h after Ba-wt LPS or PBS injection of C57BL/6 mice. CXCL1 (magenta), gp38 (green), Ly6G (cyan). Scale bar 100 µm. 3 mice per group, n = 3. c Confocal microscopy images of wholemount staining of omenta from Ba-wt LPS- or PBS-injected C57BL/6 mice at 48 h post-stimulation. CD3 (blue), gp38 (green), CXCL1 (red), Ly6G (cyan). Dotted circles indicate the contour of the milky spot (MS). Scale bar 100 µm. 2-3 mice per group, n = 3. d Flow cytometry plots of Ly6C hi monocytes secreting CXCL1 + from omenta harvested at 48 h post-administration of C57BL/6 mice with Ba-wt LPS or PBS (PBS, pooled cells from 6 mice). e Flow cytometry of indicated cells, analyzed as in ( d ) for CXCL1 positivity, from omenta harvested at 48 h post-injection of C57BL/6 mice with Ba-wt or Ec LPS, or PBS. Each symbol represents one animal. Mean from pooled data of normalized absolute numbers of CXCL1 + cells ± SD. 3-4 mice per group, n = 2. Significant differences are shown (Multiple comparison Kruskal-Wallis test, followed by post-hoc Dunn’s test). Absence of p -value, non-significant. f Spectral flow analysis of Ba-infected (red) omenta at Day 8 p.i. showing representative CXCL1 expression profiles of the myeloid cells indicated, together with their fluorescence minus one (FMO) control (top), and normalized absolute numbers of these omental CXCL1 + Ba-infected and PBS control (black) cells (bottom). Each symbol represents one animal. 1-2 mice per group, n = 4. Mean ± SD. Significant differences are shown (Mann-Whitney test, two-sided). Absence of p -value, non-significant. Source data are provided as a Source Data file.

Article Snippet: The dosage of murine CXCL1 (Cusabio, CSBE17286m) or IL-1RA (Thermofisher, EMIL1RN) was done by ELISA following the manufacturer’s instructions.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Confocal Microscopy, Staining, Flow Cytometry, Comparison, Infection, Expressing, Fluorescence, Control, MANN-WHITNEY