Journal: International immunopharmacology

Article Title: Sophora subprostrate polysaccharide targets LncRNA MSTRG.5823.1 to suppress PCV2-mediated immunosuppression via TNF/NF-κB signaling.

doi: 10.1016/j.intimp.2024.112701

Figure Lengend Snippet: Fig. 4. Validation of differentially expressed mRNAs, lncRNAs, and miRNAs (n ¼ 3). Notes: (A) mRNA expression levels of Ccl4, Clec4e, CXCL2, Acod1, Bcl2a1d, Ccl3, Dusp2, Dusp5, Glrx, IL27, Itga5, Nfkbia, Nfkbid, Nfkbiz, Nlrp3, Pdgfb, Tnf and Tnfrsf1b. (B) lncRNA expression levels of ENSMUST00000174829, ENSMUST0000034789, ENSMUST0000074287, ENSMUST0000022589, MSTRG.12908.1 and MSTRG.5823.1. (C) miRNA expression levels of miR-21-z, miR-17-x, miR-212-x, miR-217-x, and miR-342-y.

Article Snippet: The primary antibody against rabbit CXCL2 (pb0900, 1:500 dilution) was obtained from BOSTER (California, USA).

Techniques: Biomarker Discovery, Expressing

Journal: International immunopharmacology

Article Title: Sophora subprostrate polysaccharide targets LncRNA MSTRG.5823.1 to suppress PCV2-mediated immunosuppression via TNF/NF-κB signaling.

doi: 10.1016/j.intimp.2024.112701

Figure Lengend Snippet: Fig. 5. CXCL2-miR-217-X-MSTRG.5823.1 network construction. Notes: (A) Co differentially expressed mRNA, miRNA and lncRNA in the ctrl vs PCV2 com- parison group and PCV2 vs PCV2 + SSP comparison group shown by Venn diagram. (B) KEGG pathway analysis of 78 differentially co-expressed mRNAs (C) GO pathway analysis of 78 differentially co-expressed mRNAs. (D) Subcellular localization of lncRNA MSTRG.5823.1, with MSTRG.5823.1 probe in red and DAPI nuclear staining in blue, and no overlap between red and blue, indicating that MSTRG.5823.1 is located in the cytoplasm. (E) Prediction of targeting relationship between MSTRG.5823.1-miR-217-x- CXCL2. (F) In the ceRNA network, red represents up-regulation, green represents down-regulation, circles represent mRNA, triangles represent miRNAs, diamonds represent lncRNAs, and the solid line indicates the targeting relationship. (G) Targeted binding relationship between CXCL2 and miR- 217-x (n = 3). (H) Target binding relationship between MSTRG.5823.1 and miR-217-x (n = 3). Error bars are represented as means ± SD. *p value < 0.05. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The primary antibody against rabbit CXCL2 (pb0900, 1:500 dilution) was obtained from BOSTER (California, USA).

Techniques: Comparison, Staining, Binding Assay

Journal: International immunopharmacology

Article Title: Sophora subprostrate polysaccharide targets LncRNA MSTRG.5823.1 to suppress PCV2-mediated immunosuppression via TNF/NF-κB signaling.

doi: 10.1016/j.intimp.2024.112701

Figure Lengend Snippet: Fig. 6. Effect of miR-217-x in PCV2-infected RAW264.7 cells (n ¼ 3). Notes: (A) Effect of overexpression miR-217-x on the expression levels of miR-217-x, MSTRG.5823.1, Rel and TNF-α in PCV2 infected RAW264.7 cells. (B) Effect of overexpression miR-217-x on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in PCV2 infection group. (C-E) Effects of overexpression miR-217-x on the expression levels of CXCl2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65 and p-p65 proteins in PCV2 infected RAW264.7 cells. (F) Effect of knockdown miR-217-x on the expression levels of miR-217-x, MSTRG.5823.1, Rel and TNF-α. (G) Effect of knockdown miR-217-x on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in PCV2 infected RAW264.7 cells. (H-J) Effect of knockdown miR-217-x on the expression levels of CXCL2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65 and p-p65 proteins in PCV2 infected RAW264.7 cells. Error bars are represented as means ± SD. Shoulder labels with the same alphabet indicate non-significant differences (p > 0.05), while labels with different alphabets indicate significant differences (p < 0.05).

Article Snippet: The primary antibody against rabbit CXCL2 (pb0900, 1:500 dilution) was obtained from BOSTER (California, USA).

Techniques: Infection, Over Expression, Expressing, Knockdown

Journal: International immunopharmacology

Article Title: Sophora subprostrate polysaccharide targets LncRNA MSTRG.5823.1 to suppress PCV2-mediated immunosuppression via TNF/NF-κB signaling.

doi: 10.1016/j.intimp.2024.112701

Figure Lengend Snippet: Fig. 7. The function of miR-217-x in SSP-treated PCV2-infected RAW264.7 cells (n ¼ 3). Notes: (A) Effect of overexpression miR-217-x on the expression levels of miR-217-x, MSTRG.5823.1, Rel and TNF-α in SSP treatment group. (B) Effect of overexpression miR-217-x on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in SSP-treated −RAW264.7 cells infected with PCV2. (C-E) Effect of overexpression miR-217-x on the expression levels of CXCL2, TNF-α, IKB, p-IKB, IKKβ, p- IKKβ, p65and p-p65 proteins in SSP treatment group. (F) Effect of knockdown miR-217-x on the expression levels of miR-217-x, MSTRG.5823.1, Rel and TNF-α in SSP-treated RAW264.7 cells infected with PCV2. (G) Effect of knockdown miR-217-x on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in SSP treatment group. (H-J) Effect of knockdown miR-217-x on the protein expression levels of CXCL2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65and p-p65 in SSP treatment group. Error bars are represented as means ± SD Shoulder labels with the same alphabet indicate non-significant differences (p > 0.05), while labels with different alphabets indicate significant differences (p < 0.05).

Article Snippet: The primary antibody against rabbit CXCL2 (pb0900, 1:500 dilution) was obtained from BOSTER (California, USA).

Techniques: Infection, Over Expression, Expressing, Knockdown

Journal: International immunopharmacology

Article Title: Sophora subprostrate polysaccharide targets LncRNA MSTRG.5823.1 to suppress PCV2-mediated immunosuppression via TNF/NF-κB signaling.

doi: 10.1016/j.intimp.2024.112701

Figure Lengend Snippet: Fig. 8. The function of lncRNA-MSTRG.5823.1 in PCV2-infected RAW264.7 cells (n ¼ 3). Notes: (A) Effect of overexpression of MSTRG.5823.1 on the expression levels of miR-217-x, MSTRG.5823.1, Rel and TNF-α in PCV2-infected RAW264.7 cells. (B) Effect of overexpression of MSTRG.5823.1 on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in PCV2-infected RAW264.7 cells. (C-E) Effect of overexpression of MSTRG.5823.1 on the protein expression levels of CXCL2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65and p-p65 in PCV2-infected RAW264.7 cells. (F) Effect of knockdown MSTRG.5823.1 on the expression levels of miR- 217-x, MSTRG.5823.1, Rel and TNF-α. (G) Effect of knockdown MSTRG.5823.1 on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in PCV2-infected RAW264.7 cells. (H-J) Effect of knockdown MSTRG.5823.1 on the protein expression levels of CXCL2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65 and p-p65 in PCV2- infected RAW264.7 cells. Error bars are represented as means ± SD. Shoulder labels with the same alphabet indicate non-significant differences (p > 0.05), while labels with different alphabets indicate significant differences (p < 0.05).

Article Snippet: The primary antibody against rabbit CXCL2 (pb0900, 1:500 dilution) was obtained from BOSTER (California, USA).

Techniques: Infection, Over Expression, Expressing, Knockdown

Journal: International immunopharmacology

Article Title: Sophora subprostrate polysaccharide targets LncRNA MSTRG.5823.1 to suppress PCV2-mediated immunosuppression via TNF/NF-κB signaling.

doi: 10.1016/j.intimp.2024.112701

Figure Lengend Snippet: Fig. 9. The function of lncRNA-MSTRG.5823.1 in SSP-treated PCV2-infected RAW264.7 cells (n ¼ 3). Notes: (A) Effect of overexpression of MSTRG.5823.1 on the expression levels of miR-217-x, MSTRG.5823.1, Rel, TNF-α in SSP treatment group. (B) Effect of overexpression of MSTRG.5823.1 on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in SSP treatment group. (C-E) Effect of overexpression of MSTRG.5823.1 on the protein expression levels of CXCL2, TNF-α, IKB, p- IKB, IKKβ, p-IKKβ, p65 and p-p65 in SSP treatment group. (F) Effect of knockdown MSTRG.5823.1 on the expression levels of miR-217-x, MSTRG.5823.1, Rel and TNF-α in SSP treatment group. (G) Effect of knockdown MSTRG.5823.1 on the expression levels of CXCL2, CXCL10, IL-1β mRNA in SSP treatment group. (H-J) Effect of knockdown MSTRG.5823.1 on the expression levels of CXCL2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65 and p-p65 proteins in SSP treatment group. Error bars are represented as means ± SD. Shoulder labels with the same alphabet indicate non-significant differences (p > 0.05), and shoulder labels with different alphabets indicate significant differences (p < 0.05).

Article Snippet: The primary antibody against rabbit CXCL2 (pb0900, 1:500 dilution) was obtained from BOSTER (California, USA).

Techniques: Infection, Over Expression, Expressing, Knockdown

Journal: International immunopharmacology

Article Title: Sophora subprostrate polysaccharide targets LncRNA MSTRG.5823.1 to suppress PCV2-mediated immunosuppression via TNF/NF-κB signaling.

doi: 10.1016/j.intimp.2024.112701

Figure Lengend Snippet: Fig. 10. Effect of the MSTRG.5823.1/miR-217-x/CXCL2 axis on immunosuppression in PCV2-infected RAW264.7 cells (n ¼ 3). Notes: (A) Effect of over- expression of miR-217-x and MSTRG.5823.1 on the expression levels of miR-217-x, MSTRG.5823.1, Rel and TNF-α in PCV2 infection group. (B) Effect of over- expression of miR-217-x and MSTRG.5823.1 on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in in PCV2 infection group. (C-E) Effect of overexpression of miR-217-x and MSTRG.5823.1 on CXCL2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65 and p-p65 protein expression levels in in PCV2 infection group. (F) Effect of knockdown miR-217-x and MSTRG.5823.1 on the expression levels of miR-217-x, MSTRG.5823.1, Rel and TNF-α. (G) Effect of knockdown miR-217-x and MSTRG.5823.1 on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in PCV2-infected RAW264.7 cells. (H-J) Effect of knockdown miR-217-x and MSTRG.5823.1 on the protein expression levels of CXCL2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65 and p-p65 in PCV2-infected RAW264.7 cells. Error bars are rep- resented as means ± SD. Shoulder labels with the same alphabet indicate non-significant differences (p > 0.05), while labels with different alphabets indicate significant differences (p < 0.05).

Article Snippet: The primary antibody against rabbit CXCL2 (pb0900, 1:500 dilution) was obtained from BOSTER (California, USA).

Techniques: Infection, Over Expression, Expressing, Knockdown

Journal: International immunopharmacology

Article Title: Sophora subprostrate polysaccharide targets LncRNA MSTRG.5823.1 to suppress PCV2-mediated immunosuppression via TNF/NF-κB signaling.

doi: 10.1016/j.intimp.2024.112701

Figure Lengend Snippet: Fig. 11. SSP promoted activation of the TNF/NF-κB pathway by regulating MSTRG.5823.1/miR-217-x/CXCL2 axis in PCV2- infected RAW264.7 cells (n ¼ 3). Notes: (A) Effect of overexpression of miR-217-x and MSTRG.5823.1 on the expression levels of miR-217-x, MSTRG.5823.1, Rel and TNF-α in SSP treatment group. (B) Effect of overexpression of miR-217-x and MSTRG.5823.1 on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in SSP treatment group. (C-E) Effect of overexpression of miR-217-x and MSTRG.5823.1 on the expression levels of CXCL2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65and p-p65 proteins in SSP treatment group. (F) Effect of knockdown miR-217-x and MSTRG.5823.1 on the expression levels of miR-217-x, MSTRG.5823.1 and Rel, TNF-α in SSP treatment group. (G) Effect of knockdown miR-217-x and MSTRG.5823.1 on the expression levels of CXCL2, CXCL10, and IL-1β mRNA in SSP treatment group. (H-J) Effect of knockdown miR-217-x and MSTRG.5823.1 on CXCL2, TNF-α, IKB, p-IKB, IKKβ, p-IKKβ, p65 and p-p65 protein expression levels in SSP treatment group. Error bars are represented as means ± SD. Shoulder labels with the same alphabet indicate non-significant differences (p > 0.05), while labels with different alphabets indicate significant differences (p < 0.05).

Article Snippet: The primary antibody against rabbit CXCL2 (pb0900, 1:500 dilution) was obtained from BOSTER (California, USA).

Techniques: Activation Assay, Infection, Over Expression, Expressing, Knockdown

Journal: International immunopharmacology

Article Title: Sophora subprostrate polysaccharide targets LncRNA MSTRG.5823.1 to suppress PCV2-mediated immunosuppression via TNF/NF-κB signaling.

doi: 10.1016/j.intimp.2024.112701

Figure Lengend Snippet: Fig. 12. Effects of SSP on the regulation of PCV2-induced immunosuppression model in mice (n ¼ 12). Notes: (A) Schematic representation of PCV2-induced immunosuppression in vivo model: BALB/c mice were injected intraperitoneally with PCV2 (TCID50 = 10-5/0.1 mL) on the first day of the experiment, and SSP (200 mg/kg•BW) was administered by gavage every day for 7 days. (B-E) secretion levels of CXCL2, CXCL10, TNF-α and IL-1β in thymus tissue homogenate. (F) His- topathological examination of lungs, spleens, livers, and thymus of all three animal groups. [control group, PCV2-infected group, PCV2 + SSP (200 mg/kg•BW)]. scale Bar: 100 µm. (Gray boxes indicate the site of PCV2-induced lesions in tissues. scale Bar: 20 µm.) (G-J) Protein expression levels of TNF-α and CXCL2 in lungs, spleens, livers, and thymuses of all three groups of animals [control group, PCV2-infected group, PCV2 + SSP (200 mg/kg•BW)]. scale Bar: 100 µm. (K) Expression levels of p65 and p-p65 in the thymus from all three animal groups[control group, PCV2-infected group, PCV2 + SSP (200 mg/kg•BW)]. scale Bar: 50 µm. Error bars are represented as means ± SE shoulder labels with the same alphabet indicate non-significant differences (p > 0.05), while labels with different alphabets indicate significant differences (p < 0.05).

Article Snippet: The primary antibody against rabbit CXCL2 (pb0900, 1:500 dilution) was obtained from BOSTER (California, USA).

Techniques: In Vivo, Injection, Control, Infection, Expressing

Journal: ImmunoTargets and Therapy

Article Title: NLRP3 Inflammasome Upregulates PD-L1 in Ovarian Cancer and Contributes to an Immunosuppressive Microenvironment

doi: 10.2147/ITT.S495564

Figure Lengend Snippet: Inhibition of the NLRP3 inflammasome leads to a decrease in immunosuppressive cells in murine ovarian cancer. ( a – c ) Flow cytometry analysis of PD-1 + CD4 + T cells ( a ), effector CD8 + ( b ) and CD4 + ( c ) T cells in spleens and tumors of mice treated with PBS (as control) or MCC950, and their quantification and statistical evaluation. ( d – f ) Representative flow cytometry graphs (left panel) and bar charts (right panel) demonstrating the frequencies and statistical evaluation of MDSCs ( d ), TAMs ( e ), and Tregs ( f ) in the tumor and spleen of control and MCC950-treated mice. ( g ) Analysis of CXCL2 protein expression in murine tumor tissues under different treatment conditions with and without MCC950 using Western blot. (All quantitative data are presented as mean ± SD from at least three biologically independent experiments, ns, no significance. **, p < 0.01, ***, p < 0.001, ****, p < 0.0001). “#” represents each independent experiment.

Article Snippet: The following antibodies were utilized in this study: Rabbit Polyclonal anti-NLRP3 (CY5651), Rabbit Monoclonal anti- PD-L1 (CD274) Antibody (Abways, Shanghai, China; CY5980-20); Rabbit Polyclonal anti- Cleaved-Caspase-1(p20) (YC0022-20), Rabbit Polyclonal anti- IL-1β (YT5201), Rabbit Polyclonal anti- CXCL2(YT7842-20) (ImmunoWay Biotechnology, Plano, TX, USA).

Techniques: Inhibition, Flow Cytometry, Control, Expressing, Western Blot