Journal: CytoJournal

Article Title: Tumor-associated macrophage-derived chemokine ligand 5 regulates the Hypoxia-inducible factor-1α/vascular endothelial growth factor signaling pathway to promote breast cancer invasion and migration

doi: 10.25259/Cytojournal_6_2025

Figure Lengend Snippet: Analysis of CCL5 expression and clinical relevance in breast cancer. (a) CCL5 expression in breast invasive carcinoma versus normal breast tissues analyzed through gene expression profiling interactive analysis database ( http://gepia.cancer-pku.cn/index.html ); n (T) = 1085, n (N) = 291. (b) Kaplan–Meier overall survival curves comparing patients with BC with high and low CCL5 expression; n (high) = 535, n (low) = 535. (c) Violin plot showing CCL5 expression across different TNM stages of BC. (d) ELISA-based quantification of CCL5 secretion in MCF-10 and ZR-75-30 cells; n = 3. ✶ P < 0.05, ✶ ✶ ✶ P < 0.001. CCL5: Chemokine ligand 5; BC: Breast cancer, ELISA: Enzyme-linked immunosorbent assay.

Article Snippet: After each group of cells was treated, the cell culture supernatant was collected, and the CCL5 content was detected using a CCL5 ELISA kit (HB2330-Hu, Shanghai Hengyuan Biological Co., Ltd., Shanghai, China).

Techniques: Expressing, Gene Expression, Enzyme-linked Immunosorbent Assay

Journal: CytoJournal

Article Title: Tumor-associated macrophage-derived chemokine ligand 5 regulates the Hypoxia-inducible factor-1α/vascular endothelial growth factor signaling pathway to promote breast cancer invasion and migration

doi: 10.25259/Cytojournal_6_2025

Figure Lengend Snippet: Effects of 20 or 40 ng/mL CCL5 on proliferation, migration, invasion, and EMT-related protein expression in ZR-75-30 cells. (a) Cell viability evaluated by CCK-8 assay after treatment with different concentrations of CCL5. (b) Wound healing assay for assessing cell migration at 0 and 24 h (Scale bar: 50 μm, ×20). (c) Transwell assay for assessing cell invasion under varying CCL5 concentrations (Scale bar: 50 μm, ×20). (d-h) Western blot analysis and quantification of EMT markers (E-cadherin and vimentin) and metastasis-associated proteins (MMP2 and MMP9). ✶ P < 0.05, ✶ ✶ P < 0.01, ✶ ✶ ✶ P < 0.001; n = 3. CCL5: Chemokine ligand 5; EMT: Epithelial-mesenchymal transition, MMP2: Matrix metalloproteinase-2, MMP9: Matrix metalloproteinase-9.

Article Snippet: After each group of cells was treated, the cell culture supernatant was collected, and the CCL5 content was detected using a CCL5 ELISA kit (HB2330-Hu, Shanghai Hengyuan Biological Co., Ltd., Shanghai, China).

Techniques: Migration, Expressing, CCK-8 Assay, Wound Healing Assay, Transwell Assay, Western Blot

Journal: CytoJournal

Article Title: Tumor-associated macrophage-derived chemokine ligand 5 regulates the Hypoxia-inducible factor-1α/vascular endothelial growth factor signaling pathway to promote breast cancer invasion and migration

doi: 10.25259/Cytojournal_6_2025

Figure Lengend Snippet: Effects of TAM-derived CCL5 on ZR-75-30 cell proliferation, migration, and invasion. (a) CCL5 secretion in MCF-10, ZR-75-30, M0, and M2-polarized macrophages assessed by ELISA. (b and c) Confirmation of CCL5 knockdown efficiency in TAMs by Western blot and RT-qPCR. (d) Cell viability of ZR-75-30 cells following co-culture with different TAM-conditioned media, as assessed by CCK-8 assay. (e) Cell migration evaluated by wound healing assay (Scale bar: 50 μm, ×20). (f) Invasive capacity analyzed using Transwell assay (Scale bar: 50 μm, ×20). ✶ P < 0.05, ✶ ✶ P < 0.01, and ✶ ✶ ✶ P < 0.001; n = 3. CCL5: Chemokine ligand 5; TAM: Tumor-associated macrophage, ELISA: Enzyme-linked immunosorbent assay.

Article Snippet: After each group of cells was treated, the cell culture supernatant was collected, and the CCL5 content was detected using a CCL5 ELISA kit (HB2330-Hu, Shanghai Hengyuan Biological Co., Ltd., Shanghai, China).

Techniques: Derivative Assay, Migration, Enzyme-linked Immunosorbent Assay, Knockdown, Western Blot, Quantitative RT-PCR, Co-Culture Assay, CCK-8 Assay, Wound Healing Assay, Transwell Assay

Journal: CytoJournal

Article Title: Tumor-associated macrophage-derived chemokine ligand 5 regulates the Hypoxia-inducible factor-1α/vascular endothelial growth factor signaling pathway to promote breast cancer invasion and migration

doi: 10.25259/Cytojournal_6_2025

Figure Lengend Snippet: Effect of TAM-derived CCL5 on EMT- and invasion-related protein expression in ZR-75-30 cells. (a) Representative Western blot images of E-cadherin, vimentin, MMP2, and MMP9 following treatment with different TAM-conditioned media. (b-e) Quantification of protein expression levels normalized to GAPDH. ✶ P < 0.05, ✶ ✶ P < 0.01, and ✶ ✶ ✶ P < 0.001; n = 3. CCL5: Chemokine ligand 5; TAM: Tumor-associated macrophage, EMT: Epithelial-mesenchymal transition.

Article Snippet: After each group of cells was treated, the cell culture supernatant was collected, and the CCL5 content was detected using a CCL5 ELISA kit (HB2330-Hu, Shanghai Hengyuan Biological Co., Ltd., Shanghai, China).

Techniques: Derivative Assay, Expressing, Western Blot

Journal: CytoJournal

Article Title: Tumor-associated macrophage-derived chemokine ligand 5 regulates the Hypoxia-inducible factor-1α/vascular endothelial growth factor signaling pathway to promote breast cancer invasion and migration

doi: 10.25259/Cytojournal_6_2025

Figure Lengend Snippet: Effects of TAM-derived CCL5 and HIF-1α on proliferation, migration, and invasion of ZR-75-30 cells. (a and b) mRNA expression levels of HIF-1α and CCL5 in ZR-75-30 cells co-cultured with TAMs transduced with sh-CCL5 or OE-HIF-1α, as assessed by RT-qPCR. (c-e) Western blot and quantification of HIF-1α and VEGF protein levels. (f) Cell viability measured by CCK-8 assay. (g) Migration assessed by wound healing assay (Scale bar: 50 μm, ×20). (h) Cell invasion evaluated using Transwell assay (Scale bar: 50 μm, ×20). ✶ P < 0.05, ✶ ✶ P < 0.01, and ✶ ✶ ✶ P < 0.001; n = 3. TAM: Tumor-associated macrophage, CCL5: Chemokine ligand 5, HIF-1α: Hypoxia-inducible factor-1α, VEGF: Vascular endothelial growth factor.

Article Snippet: After each group of cells was treated, the cell culture supernatant was collected, and the CCL5 content was detected using a CCL5 ELISA kit (HB2330-Hu, Shanghai Hengyuan Biological Co., Ltd., Shanghai, China).

Techniques: Derivative Assay, Migration, Expressing, Cell Culture, Transduction, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Wound Healing Assay, Transwell Assay

Journal: CytoJournal

Article Title: Tumor-associated macrophage-derived chemokine ligand 5 regulates the Hypoxia-inducible factor-1α/vascular endothelial growth factor signaling pathway to promote breast cancer invasion and migration

doi: 10.25259/Cytojournal_6_2025

Figure Lengend Snippet: Regulation of EMT and angiogenesis in ZR-75-30 cells by TAM-derived CCL5 via HIF-1α/VEGF pathway. (a) Western blot analysis of EMT-related (E-cadherin and vimentin) and metastasis-related (MMP2 and MMP9) protein levels in ZR-75-30 cells co-cultured with differentially modified TAMs. (b-e) Quantification of protein expression normalized to GAPDH. (f) Angiogenic activity assessed by HUVEC tube formation assay (Scale bar: 50 μm, ×20). ✶ P < 0.05, ✶ ✶ P < 0.01, and ✶ ✶ ✶ P < 0.001; n = 3. TAM: Tumor-associated macrophage, CCL5: Chemokine ligand 5, EMT: Epithelial-mesenchymal transition, HUVEC: Human umbilical vein endothelial cells.

Article Snippet: After each group of cells was treated, the cell culture supernatant was collected, and the CCL5 content was detected using a CCL5 ELISA kit (HB2330-Hu, Shanghai Hengyuan Biological Co., Ltd., Shanghai, China).

Techniques: Derivative Assay, Western Blot, Cell Culture, Modification, Expressing, Activity Assay, HUVEC Tube Formation Assay

Journal: iScience

Article Title: Serglycin modulates inflammation and metabolism in macrophages

doi: 10.1016/j.isci.2026.115235

Figure Lengend Snippet: SRGN deficiency reshapes the inflammatory secretome and transcriptional programs in human macrophages (A) Volcano plot of differentially secreted proteins between SRGN −/− and wild-type THP-1 macrophages under M1 polarization. Among 1,507 quantified proteins, 53 were significantly altered (adjusted p < 0.05), with serglycin being the most downregulated protein in knockout cells. (B) Validation of selected targets by RT-qPCR. SRGN −/− M1 macrophages showed significantly increased expression of IL6 and TNF and reduced expression of CCL5 compared with wild-type cells (mean ± SEM; unpaired Student’s t test; p < 0.05, ∗ p < 0.01, and ∗∗∗ p < 0.0001; n = 4). (C) ELISA quantification of secreted cytokines in culture supernatants. SRGN −/− macrophages secreted significantly less TNF-α, CCL5, and IL-6 compared with wild-type macrophages ( n = 4), consistent with proteomics and RNA-seq data. (D) Transmission electron microscopy (TEM) images of THP-1 M0 and M1 macrophages. Scale bars, 5 μm. Vesicles were manually annotated and quantified in 10 cells per experimental group. The number of vesicles per cell and the percentage of cellular area occupied by vesicles were significantly reduced in both M0 and M1 SRGN −/− macrophages compared with wild-type macrophages. (E) Phagocytosis assay using fluorescently labeled bioparticles. SRGN −/− macrophages exhibited reduced phagocytic capacity under both M0 and M1 conditions (mean ± SEM; unpaired Student’s t test; p < 0.05 and ∗∗ p < 0.001; n = 6).

Article Snippet: THP-1: cytokine levels were measured using ELISA kits from R&D Systems: CCL5 (Cat. No. DY278), IL6 (Cat. No. DY206), IL-1β (Cat. No. DY201), TGF-β (Cat. No. DY240; activation kit DY010), and TNF-α (Cat. No. DY210), following the manufacturer’s instructions.

Techniques: Knock-Out, Biomarker Discovery, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Transmission Assay, Electron Microscopy, Phagocytosis Assay, Labeling