Journal: Journal of Advanced Research

Article Title: The pro-cancer and immunosuppressive activity of macrophage-transformed cancer-associated fibroblasts in oral squamous cell carcinoma

doi: 10.1016/j.jare.2025.07.027

Figure Lengend Snippet: MMTs recruit a large number of MDSCs through CXCL10 pathway. A Significantly upregulated genes in MMTs versus non-MMT fibroblasts from OSCC database (GSA-Human: HRA007439). B Differences in CXCL10 expression between non-MMT fibroblasts and MMTs were demonstrated in HNSCC (figure S1). C CXCL10 expression and release were detected by PCR and ELISA. D-H Exogenous CXCL10 was added to BMDMs, while the CXCL10 inhibitor AMG487 was administered to MMTs, followed by co-culturing with MDSCs. The migration of MDSCs was evaluated using transwell chambers and crystal violet staining ( D ). Flow cytometry was used to detect cell apoptosis ( E ), macrophage markers (F4/80 + CD11b + ) ( F ), as well as CFSE for assessing T cell proliferation inhibition ( G ). PCR and ELISA were employed to measure the expression of immunosuppressive factors IL10 and iNOS ( H ). I, J Tumor tissue photographs ( I ) and the tumor growth curve were taken and subjected to statistical analysis regarding volume and weight ( J ). K-M Flow cytometry was used to detect the proportion of MDSCs in the spleen ( K ) and tumors ( L ) of each group, while immunofluorescence staining ( M ) revealed the densest field of MDSCs within the tumor tissues across all groups. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs experimental group. “ns” represents no statistical significance. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: According to the manufacturer's instructions, the cytokine concentrations of TGFβ1, iNOS, IL-10, and CXCL10 in the supernatant were determined using ELISA kits (Boster, California, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Migration, Staining, Flow Cytometry, Inhibition, Immunofluorescence

Journal: Clinical and Experimental Medicine

Article Title: CXCL12-mediated T cell infiltration drives breast cancer metastasis

doi: 10.1007/s10238-026-02126-2

Figure Lengend Snippet: The differences in the expression and clinical correlation of CXCL10 were evaluated in breast cancer patients. ( A ) The role of CXCL10 in breast cancer was predicted with the CanerSEA database. ( B ) Differences in the expression of CXCL10 in different types of breast cancer were analyzed with the TIMER2.0 database. ( C ) Based on GEPIA2 breast cancer data, a volcano map was drawn with bioinformatics to determine the expression level of CXCL10. ( D ) The relationship between CXCL10 and the clinical prognosis of breast cancer patients was analyzed with the TIMER2.0 database. ( E ) The expression level of CXCL10 in adjacent and cancerous tissues was detected by immunohistochemistry. ( F ) The expression differences in different clinical stages were analyzed with the GEPIA2 database. (*** p < 0.001)

Article Snippet: After 24 h of incubation, the culture supernatants were analyzed with a human CXCL10 ELISA kit (R&D Systems, QK266).

Techniques: Expressing, Immunohistochemistry

Journal: Clinical and Experimental Medicine

Article Title: CXCL12-mediated T cell infiltration drives breast cancer metastasis

doi: 10.1007/s10238-026-02126-2

Figure Lengend Snippet: The composition of cells in the breast cancer tissue microenvironment, interactions between cells, and differences in CXCL10 expression between breast cancer cells and T cells were analyzed with the single-cell sequencing data. ( A - B ) The composition of the main cells in the microenvironment of breast cancer tissue was analyzed with the NGDC database. ( C ) The interaction relationships between various types of cells were analyzed. ( D ) The CXCL10 expression levels were analyzed in various types of cells

Article Snippet: After 24 h of incubation, the culture supernatants were analyzed with a human CXCL10 ELISA kit (R&D Systems, QK266).

Techniques: Expressing, Single Cell, Sequencing

Journal: Clinical and Experimental Medicine

Article Title: CXCL12-mediated T cell infiltration drives breast cancer metastasis

doi: 10.1007/s10238-026-02126-2

Figure Lengend Snippet: Correlation and differential expression analyses of CXCL10-interacting molecules were performed in BC. ( A ) CXCL10-interacting molecules were analyzed with the String database. ( B - E ) The correlations between CXCL10 and CCL13, CXCL6, PF4V1, and CXCL12 were analyzed with the GEPIA2 database. ( F - I ) Differences in the expression of CCL13, CXCL6, PF4V1 and CXCL12 in breast cancer were analyzed with the GEPIA2 database. ( J ) The differential expression of CXCL10 and CXCL12 in breast epithelial cells and breast cancer cells was detected by western blotting. (* p < 0.05)

Article Snippet: After 24 h of incubation, the culture supernatants were analyzed with a human CXCL10 ELISA kit (R&D Systems, QK266).

Techniques: Quantitative Proteomics, Expressing, Western Blot

Journal: Clinical and Experimental Medicine

Article Title: CXCL12-mediated T cell infiltration drives breast cancer metastasis

doi: 10.1007/s10238-026-02126-2

Figure Lengend Snippet: The differences in the expression and clinical correlation of CXCL12 were evaluated in BC patients. ( A ) Differences in CXCL12 expression in different types of breast cancer were analyzed with the TIMER2.0 database. ( B ) Based on GEPIA2 breast cancer data, a volcano map was drawn by bioinformatics to determine the expression of CXCL12. ( C ) Differences in the expression of CXCL12 in different clinical stages were analyzed with the GEPIA2 database. ( D ) The relationship between CXCL12 and the prognosis of breast cancer patients was analyzed with the TIMER2.0 database. ( E - F ) The expression of CXCL12 in different types of cells was analyzed with single-cell sequencing from the NGDC database. ( G ) The expression level of CXCL10 in adjacent and cancerous tissues was detected by immunohistochemistry. (*** p < 0.001)

Article Snippet: After 24 h of incubation, the culture supernatants were analyzed with a human CXCL10 ELISA kit (R&D Systems, QK266).

Techniques: Expressing, Single Cell, Sequencing, Immunohistochemistry

Journal: Clinical and Experimental Medicine

Article Title: CXCL12-mediated T cell infiltration drives breast cancer metastasis

doi: 10.1007/s10238-026-02126-2

Figure Lengend Snippet: The impact of CXCL12 on breast cancer and its correlation with CXCL10 were evaluated. ( A ) CXCL12 knockdown efficiency was detected by Q-PCR. ( B - C ) The impact of CXCL12 knockdown on cell invasion ability was detected by transwell assays. ( D - E ) The effect of CXCL12 knockdown on cell migration ability was detected by scratch assays. ( F ) The effect of CXCL12 knockdown on CXCL10 expression was detected by Q-PCR. ( G ) The effect of CXCL12 knockdown on CXCL10 secretion levels was detected by ELISA. (*** p < 0.001)

Article Snippet: After 24 h of incubation, the culture supernatants were analyzed with a human CXCL10 ELISA kit (R&D Systems, QK266).

Techniques: Knockdown, Migration, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Clinical and Experimental Medicine

Article Title: CXCL12-mediated T cell infiltration drives breast cancer metastasis

doi: 10.1007/s10238-026-02126-2

Figure Lengend Snippet: Correlations between CXCL12 and CXCL10 and CD4 + T and CD8 + T cells in the inflammatory microenvironment of breast cancer patients were evaluated, and correlations with the clinical prognosis of breast cancer patients were evaluated. ( A ) The role of CXCL10 in breast cancer was predicted with the CanerSEA database. ( B ) The expression levels of CXCL10 and CXCL12 in different types of T lymphocytes were analyzed in different types of breast cancer with the TIMER2.0 database. ( C - F ) The clinical prognosis under different conditions was analyzed with the TIMER2.0 database. ( G ) CD4 + T cells and CD8 + T cells in breast cancer tissues were analyzed by immunofluorescence

Article Snippet: After 24 h of incubation, the culture supernatants were analyzed with a human CXCL10 ELISA kit (R&D Systems, QK266).

Techniques: Expressing, Immunofluorescence

Journal: bioRxiv

Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling

doi: 10.64898/2026.03.24.712780

Figure Lengend Snippet: ( A ) HEK293T reporter cells were treated with 6.0 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle, cells were imaged by confocal microscopy at 6 h post treatment to examine conjugate colocalization with target TBK1 (representative of N = 3 biological replicates). Scale bar is 10 μm. ( B ) HEK293T reporter cells were treated with 8.3 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle, Western blot was performed at 6 h post treatment to examine TBK1 and IRF3 phosphorylation (representative of N = 3 biological replicates). ( C ) IRF3 reporter signal relative to buffer treatment for HEK293T reporter cells pretreated for 6 h with TBK1 inhibitor MRT67307 (TBK1i) and then treated with 8.3 μg/mL STING or Scr conjugate delivered using TransIT-X2, measured 24 h post treatment (N = 3 biological replicates). ( D ) Western blot of STING and β-actin expression in ovarian cancer cell lines KURAMOCHI and A2780. ( E-F ) KURAMOCHI and A2780 ovarian cancer cell lines were treated with 5.0 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle or 50 μM STING agonist ADU-S100. ( E ) CXCL10 and ( F ) IFN-β in supernatant was measured by ELISA 24 h post treatment (N = 3 biological replicates). Replicates where analyte was below the limit of detection (LOD) are labeled as not detected (ND), no summary statistics were computed if any replicate was ND. ( G-I ) KURAMOCHI cells were treated with 5.0 μg/mL STING or Scr conjugate using TransIT-X2 as a vehicle or 50 μM ADU-S100, mRNA sequencing was performed at 6 h post treatment (N = 4 biological replicates). ( G ) Plot of Log2 fold change of STING conjugate or ADU-S100 treatment compared to Buffer, showing high correlation between treatments. Plot of Log2 fold change of Scr conjugate or ADU-S100 treatment compared to Buffer is displayed below as a control, showing greatly reduced correlation. The coefficient of determination R 2 for line of best fit is displayed. ( H ) Gene set enrichment analysis was performed on MSigDB Hallmark gene set, normalized enrichment and adjusted P value are displayed for the 10 gene sets significantly enriched ( P < .05) when comparing STING to Scr conjugate. Normalized enrichment and adjusted P values for the same 10 gene sets are displayed for the comparison of ADU-S100 to Buffer ( I ) Heatmap of gene expression for selected genes. Replicates where a given gene was not detected are labeled ND. Data represented as geometric mean ± SD.

Article Snippet: DuoSet ELISA kits (R&D Systems) for human CXCL10 (#DY266), human IFN-β (#DY814), mouse CXCL10 (#DY466), mouse IFN-β (#DY8234), mouse IL-6 (#DY406), mouse TNF-α (#DY410), and mouse IFN-γ (#DY485) were used with 1-Step TMB ELISA Substrate Solution (Thermo Scientific) following vendor instructions.

Techniques: Confocal Microscopy, Western Blot, Phospho-proteomics, Expressing, Enzyme-linked Immunosorbent Assay, Labeling, Sequencing, Control, Comparison, Gene Expression

Journal: bioRxiv

Article Title: A multivalent peptide-polymer conjugate material mimics STING to therapeutically activate innate immune signaling

doi: 10.64898/2026.03.24.712780

Figure Lengend Snippet: ( A-B ) Mice were dosed with 20 μg of STING conjugate delivered by LNP IP and serum was collected at 0, 1, 3, 6, 10, 24, and 50 h. Serum was analyzed to measure ( A ) STING conjugate concentration by Cy5 fluorescence (showing one phase exponential decay fit to data) and ( B ) CXCL10 concentration by ELISA (N = 3 mice). Conditions where analyte was below the limit of detection (LOD) are labeled as not detected (ND). ( C ) Mice were inoculated with 3×10 6 BPPNM cells IP and dosed with 20 μg of STING or Scr conjugate (N = 3 mice) delivered by LNP IP at 14 days after inoculation. ( D ) Omental tumor, ( E ) ascites, and ( F ) serum were collected 6 h after dosing. Concentrations of CXCL10, IFN-β, IL-6, TNF-α, and IFN-γ were measured by ELISA and are reported relative to total protein concentration in tumor and ascites ( D-E ) or relative to volume in serum ( F ). Conditions where analyte was below the LOD are labeled as ND. Average fold change increases in cytokine concentration for STING conjugate treatment compared to Scr conjugate are displayed. Where cytokine level was ND, a lower bound on the fold change was computed by setting all ND replicates as the LOD. Data represented as mean ± SD.

Article Snippet: DuoSet ELISA kits (R&D Systems) for human CXCL10 (#DY266), human IFN-β (#DY814), mouse CXCL10 (#DY466), mouse IFN-β (#DY8234), mouse IL-6 (#DY406), mouse TNF-α (#DY410), and mouse IFN-γ (#DY485) were used with 1-Step TMB ELISA Substrate Solution (Thermo Scientific) following vendor instructions.

Techniques: Concentration Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Labeling, Protein Concentration