Journal: Nature Communications

Article Title: Integrated multi-omics profiling reveals neutrophil extracellular traps potentiate Aortic dissection progression

doi: 10.1038/s41467-024-55038-8

Figure Lengend Snippet: A Interaction network of main cell types in AD constructed with CellChat. B The ligand-receptor interactions between macrophages and neutrophils in AD compared to those in NA. C , D Representative images of the presence of CXCL3 + macrophages within human ( C ) and mice ( D ) dissected aorta. Scale bars = 100 μm. E , F Representative images of the presence of CXCR2 + neutrophils within human ( E ) and mice ( F ) dissected aorta. Scale bars = 100 μm. G Pearson correlation analysis showing positive linear correlation between the number of CXCL3 + macrophages and the number of CXCR2 + neutrophils within human aortic lesion ( n = 5, three fields per sample). H , I Bar plot showing the positive correlation between elastin degradation grading and CXCL3 + macrophages ( H ) and CXCR2 + neutrophils ( I ) ( n = 5, three fields per sample). Values are expressed as means ± SD. A two-sided statistical significance was set at * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Mann–Whitney U test.

Article Snippet: In the second experiment, the challenged mice were treated with either isotype control antibody ( n = 12) or CXCL3 antibody (2 mg/kg, AF5568, R&D systems, Minneapolis, MN, USA) ( n = 12) or CXCR2 antibody (2 mg/kg, MAB2164-100, R&D systems, Minneapolis, MN, USA) ( n = 12) intraperitoneally, every three days, starting at 1 week after BAPN administration.

Techniques: Construct, MANN-WHITNEY

Journal: Nature Communications

Article Title: Integrated multi-omics profiling reveals neutrophil extracellular traps potentiate Aortic dissection progression

doi: 10.1038/s41467-024-55038-8

Figure Lengend Snippet: A Schematic overview of experimental design. Mice received intraperitoneal injection of IgG anti-body, anti-CXCL3 anti-body (2 mg/kg), and anti-CXCR2 anti-body (2 mg/kg) at a fixed time once a day until death or the end point of the experiment. B The incidence of AD and aortic rupture in each group ( n = 12). Comparisons by Fisher’s exact test indicating aortic rupture rate difference among groups ( p = 0.0045; p = 0.0137; p = 0.2174; p = 0.4783). C Mice death due to aortic rupture of each indicated group ( p = 0.0408 for anti-CXCR2 vs BAPN). D – F The average of max diameter ( D ) ( p = 0.0065 for anti-CXCL3 vs Ctrl), aortic wall thickness ( E ) ( p = 0.0012 for BAPN vs Ctrl; p = 0.0284 for IgG vs Ctrl), and aortic media thickness ( F ) in each group. G Elastin degradation grading evaluation of each aorta ( p = 0.0423 for anti-CXCL3 vs BAPN; p = 0.0318 for anti-CXCR2 vs BAPN). H Representative macrographs of aortas in each group. Scale bar = 20 mm. I Representative immunohistochemistry images showing aortic dilation, false lumen formation, and elastin degradation within aortas in each group. Scale bar = 100 μm. J Representative immunofluorescence images showing NETs formation within aortas of mice. Scale bar = 100 μm. K Immunofluorescence quantification of NETs markers among each group (CitH3: p = 0.0025 for anti-CXCL3 vs BAPN; p = 0.0044 for anti-CXCR2 vs BAPN; Ly6G: p = 0.0025 for anti-CXCL3 vs BAPN; p = 0.0043 for anti-CXCR2 vs BAPN). L Immunofluorescence quantification of NETs markers within aortas with four elastin degradation grades (CitH3: p = 0.0489 for grade 2 vs grade 1; p = 0.0012 for grade 3 vs grade 1; p = 0.0473 for grade 4 vs grade 1; Ly6G: p = 0.0537 for grade 2 vs grade 1; p = 0.0425 for grade 3 vs grade 1; p = 0.0414 for grade 4 vs grade 1). Values are expressed as means ± SD. A two-sided statistical significance was set at * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Mann–Whitney U test.

Article Snippet: In the second experiment, the challenged mice were treated with either isotype control antibody ( n = 12) or CXCL3 antibody (2 mg/kg, AF5568, R&D systems, Minneapolis, MN, USA) ( n = 12) or CXCR2 antibody (2 mg/kg, MAB2164-100, R&D systems, Minneapolis, MN, USA) ( n = 12) intraperitoneally, every three days, starting at 1 week after BAPN administration.

Techniques: Injection, Immunohistochemistry, Immunofluorescence, MANN-WHITNEY

Journal: Heliyon

Article Title: hsa_circ_0007755 competitively adsorbs miR-27b-3p to mediate CXCL2 expression and recruit Th1 cells to promote hypertrophic scars development

doi: 10.1016/j.heliyon.2024.e39169

Figure Lengend Snippet: CXCL2 is the target gene of miR-27b-3p. A: starbase predicted the potential binding sites of CXCL2 and miR-27b-3p. B: Western blot tested CXCL2 in human normal skin tissue and HS tissue. C: Western blot tested CXCL2 in normal skin tissue and HS tissue of mice. D: Western blot tested CXCL2 in HFF-1 and hHSFs. E-F: Dual luciferase reporter assay and RIP analyzed the targeting relationship between CXCL2 and miR-27b-3p. G: Western blot tested the effect of overexpressed miR-27b-3p on CXCL2 expression in hHSFs. Data were expressed as mean ± SD (N = 3). ∗ P < 0.05.

Article Snippet: Primary antibodies used were: CXCL2 (MAB452, R&D Systems), Ki-67 (M7240, Dako), phospho-p65 (3033, Cell Signaling Technology), α-smooth muscle actin (α-SMA) (A5228, MilliporeSigma), collagen I (ab34710, Abcam), and GAPDH (ab8245, Abcam).

Techniques: Binding Assay, Western Blot, Luciferase, Reporter Assay, Expressing

Journal: Heliyon

Article Title: hsa_circ_0007755 competitively adsorbs miR-27b-3p to mediate CXCL2 expression and recruit Th1 cells to promote hypertrophic scars development

doi: 10.1016/j.heliyon.2024.e39169

Figure Lengend Snippet: The promotion effect of overexpressed hsa_circ_0007755 on proliferation, invasion and inflammation of hHSFs is mitigated by knockdown of CXCL2. pcDNA 3.1-hsa_circ_0007755 and si-CXCL2 were co-transfected into hHSFs. A: Western blot tested CXCL2. B: CCK-8 assayed hHSFs proliferation. C: Flow cytometry measured apoptosis rate. D: Transwell assayed invasion ability. E: Western blot analyzed Ki-67, α-SMA, collagen I, and p-p65. F: Flow cytometry detected TH1/TH2 cell changes. G: ELISA measured cytokines TNF-α, INF-γ, IL-4, and IL-10 in TH1/TH2 cells. Data were expressed as mean ± SD (N = 3). ∗ P < 0.05.

Article Snippet: Primary antibodies used were: CXCL2 (MAB452, R&D Systems), Ki-67 (M7240, Dako), phospho-p65 (3033, Cell Signaling Technology), α-smooth muscle actin (α-SMA) (A5228, MilliporeSigma), collagen I (ab34710, Abcam), and GAPDH (ab8245, Abcam).

Techniques: Knockdown, Transfection, Western Blot, CCK-8 Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Heliyon

Article Title: hsa_circ_0007755 competitively adsorbs miR-27b-3p to mediate CXCL2 expression and recruit Th1 cells to promote hypertrophic scars development

doi: 10.1016/j.heliyon.2024.e39169

Figure Lengend Snippet: Low hsa_circ_0007755 inhibits HS and regulates TH1/TH2 balance in mice. A: Western blot tested hsa_circ_0007755. B: Representative images of HE staining of mouse scar tissue. C: Flow cytometry analyzed changes in TH1/TH2 ratio in mouse blood samples. D: ELISA measured serum levels of TNF-α, INF-γ, IL-4, and IL-10. E: Western blot tested CXCL2, Ki-67, α-SMA, collagen I, and p-p65 in mouse scar tissue. Data were expressed as mean ± SD (n = 6). ∗ P < 0.05.

Article Snippet: Primary antibodies used were: CXCL2 (MAB452, R&D Systems), Ki-67 (M7240, Dako), phospho-p65 (3033, Cell Signaling Technology), α-smooth muscle actin (α-SMA) (A5228, MilliporeSigma), collagen I (ab34710, Abcam), and GAPDH (ab8245, Abcam).

Techniques: Western Blot, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Science Advances

Article Title: ACSL6-activated IL-18R1–NF-κB promotes IL-18–mediated tumor immune evasion and tumor progression

doi: 10.1126/sciadv.adp0719

Figure Lengend Snippet: ( A and B ) Kyoto Encyclopedia of Genes and Genomes (A) and gene set enrichment analysis (GSEA) analyses (B) of RNA-seq data from shNT and shACSL6 MHCC97H cells. MAPK, mitogen-activated protein kinase; FDR, false discovery rate; NES, normalized enrichment score. ( C to E ) ACSL6-depleted MHCC97H and Huh7 cells were infected with rACSL6 WT or S674A and then treated with or without IL-18 (20 ng ml −1 ) for 12 hours. The mRNA (C) and protein expression (D) levels of the indicated genes and concentrations of CXCL1 and CXCL5 (E) in the medium were detected. ( F ) ACSL6-depleted MHCC97H cells were infected with rACSL6 WT or S674A and then treated with or without IL-18 (20 ng ml −1 ) for 1 hour. Immunoblotting analyses were performed with indicated antibodies. ( G ) ChIP-qPCR analyses were performed with the indicated antibodies, and DNA was amplified with primers targeting positive sites in the GADD45B or CXCL1 gene in ACSL6-depleted MHCC97H cells with forced expression of rACSL6 WT and S674A. ( H ) Transwell assays of the migration ability of ACSL6-depleted MHCC97H or Huh7 cells with forced expression of rACSL6 WT, S674A, or S674A and MMP3. ( I ) CCK8 analyses of ACSL6-depleted MHCC97H or Huh7 cells with forced expression of rACSL6 WT, S674A, or S674A with GADD45B. * P < 0.05, ** P < 0.01, and *** P < 0.001. One-way ANOVA [(C), (E), (G), and (H)] and two-way ANOVA (I).

Article Snippet: After injection for 8 days, the tumor-bearing mice were treated by intraperitoneally injecting 100 μg of anti-CXCL1 antibody (R&D Systems, no. MAB453) and 100 μg of anti-CXCL5 antibody (Novus Biologicals, no. NBP2-22026) or IgG control twice a week for 2 weeks beginning on day 8 after subcutaneous tumor implantation.

Techniques: RNA Sequencing Assay, Infection, Expressing, Western Blot, Amplification, Migration

Journal: Science Advances

Article Title: ACSL6-activated IL-18R1–NF-κB promotes IL-18–mediated tumor immune evasion and tumor progression

doi: 10.1126/sciadv.adp0719

Figure Lengend Snippet: ( A to C ) Flow cytometry analyses of tumor-infiltrating TANs (A), TAMs (B), PD-L1 + TANs, and CD170 + TANs (C) in indicated tumors from C57BL/6 mice. ( D and E ) Acsl6-depleted Hepa1-6 cells with forced expression of rAcsl6 WT or S674A were subcutaneously injected into C57BL/6 mice, which were then treated with or without neutralizing antibodies against CXCL1 and CXCL5. Tumor volumes were recorded (D). Flow cytometry analyses of tumor-infiltrating CD8 + T cells, TANs, and TAMs (E). ( F to M ) Liver cancer mouse model was constructed by hydrodynamic transfection of AKT/NRAS/SB plasmids before the injection of AAV8-TBG-miR30-Acsl6-shRNA-GFP or AAV8-TBG-GFP-miR30-shRNA. Schematic diagram and injection timeline (F). Immunoblotting analyses were performed with indicated antibodies (G). Representative liver images are shown (H). Liver weight (I), liver weight/body weight ratio (J), and tumor numbers (K) were measured. Representative image of H&E staining of mouse liver sections are shown (L). Flow cytometry analyses of tumor-infiltrating CD8 + T cells, TANs, and TAMs in indicated tumors (M). ( N to R ) Liver cancer mouse model was generated by hydrodynamic transfection of indicated plasmids (N). Representative liver images are shown (O). Liver weights (P), liver weight/body weight ratio (Q), and tumor numbers (R) were calculated. ( S and T ) Control or Acsl6-OE Hepa1-6 cells were subcutaneously injected into C57BL/6 mice, which were then treated with or without IL-18BP. Tumor volumes (S) and tumor-infiltrating CD8 + T cells, TANs, and TAMs (T) were analyzed. [(D) and (S)] n = 5; [(F) to (R)] n = 6. * P < 0.05, ** P < 0.01, and *** P < 0.001. Student’s t test [(A) to (C), (I) to (K), (M), and (P) to (R)], two-way ANOVA [(D) and (S)], and one-way ANOVA [(E) and (T)].

Article Snippet: After injection for 8 days, the tumor-bearing mice were treated by intraperitoneally injecting 100 μg of anti-CXCL1 antibody (R&D Systems, no. MAB453) and 100 μg of anti-CXCL5 antibody (Novus Biologicals, no. NBP2-22026) or IgG control twice a week for 2 weeks beginning on day 8 after subcutaneous tumor implantation.

Techniques: Flow Cytometry, Expressing, Injection, Construct, Transfection, shRNA, Western Blot, Staining, Generated, Control

Journal: Science Advances

Article Title: ACSL6-activated IL-18R1–NF-κB promotes IL-18–mediated tumor immune evasion and tumor progression

doi: 10.1126/sciadv.adp0719

Figure Lengend Snippet: ( A to E ) Mice with established Acsl6 depletion or control Hepa1-6 tumors were treated with or without anti–PD-1. Tumor volumes (A) and survival rates (B) were recorded. Flow cytometry analyses of tumor-infiltrating CD8 + T cells (C). Measurement of the concentration of IFN-γ (D). Flow cytometry analyses of TANs and TAMs (E). ( F to J ) Acsl6 WT or S674A was reconstituted into Acsl6-depleted Hepa1-6 cells, which were then subcutaneously injected into C57BL/6 mice, and mice were subsequently treated with anti–PD-1. Tumor volumes (F) and survival rates (G) were recorded. Flow cytometry analyses of tumor-infiltrating CD8 + T cells (H). Measurement of the concentration of IFN-γ (I). Flow cytometry analyses of TANs and TAMs (J). ( K and L ) IHC staining with anti–ACSL6 pS674, anti-IκBα, and anti-CXCL1 antibodies in tumors from patients with liver cancer from ZHPH (K) and correlation analyses (L). ( M ) Kaplan-Meier analyses of overall survival according to the ACSL6 pS674 level in patients with liver cancer from ZHPH cohort. ( N ) Proposed model of the mechanism by which ACSL6 activates IL-18–NF-κB signaling to promote immune evasion and tumor progression in liver cancer. [(A) and (E)] n = 5; [(B) and (G)] n = 10. * P < 0.05, ** P < 0.01, and *** P < 0.001. Two-way ANOVA [(A) and (F)], log-rank test [(B), (G), and (M)], and one-way ANOVA [(C) to (E) and (H) to (J)].

Article Snippet: After injection for 8 days, the tumor-bearing mice were treated by intraperitoneally injecting 100 μg of anti-CXCL1 antibody (R&D Systems, no. MAB453) and 100 μg of anti-CXCL5 antibody (Novus Biologicals, no. NBP2-22026) or IgG control twice a week for 2 weeks beginning on day 8 after subcutaneous tumor implantation.

Techniques: Control, Flow Cytometry, Concentration Assay, Injection, Immunohistochemistry