antibody based protein array  (R&D Systems)

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Suppression of STK11 induces expansion of polymorphonuclear myeloid-derived suppressive cells and activation of immune signaling in breast cancer

doi: 10.1007/s00262-025-04189-8

Figure Lengend Snippet: Gene set enrichment analysis (GSEA) of immune-related pathways in Stk11-KO cells versus their parental counterparts using Kyoto Encyclopedia of Genes and Genomes (KEGG), REACTOME, and GENE Ontology (GO) gene sets. GESA was performed on RNA sequencing data from Stk11 -KO cells and their parental counterparts derived from three mouse breast cancer cell lines (PY8119, M158, and NF639). Enrichment analysis included gene sets from the KEGG, REACTOME, and GO databases. Representative enrichment plots are shown for the following pathways and biological functions: A CYTOKINE_CYTOKINE_RECEPTOR_INTERACTION. B TYPE_II_INTERFERON_TO_JAK_STAT_SIGNALING_PATHWAY. C IL6_FAMILY_TO_JAK_STAT_SIGNALING-PATHWAY. D INTERLEUKIN_10_ SIGNALING. ( E ) CHEMOKINE_RECEPTORS_BIND_CHEMOKINES. F Gene ontology biological process (GOBP)_REGULATION_OF_ACUTE_ INFLAMMATORY_RESPONSE. G Gene ontology molecular function (GOMF)_CHEMOKINE_ACTIVITY. H GOBP_POSITIVE_REGULATION_OF_ INFLAMMATORY_RESPONSE. I GOBP_POSITIVE_REGULATION_OF_ ANTIGEN_PROCESSING_AND_PRESENTATION

Article Snippet: A Mouse Cytokine Antibody Array Kit C3 (RayBiotech, Norcross, GA) was used according to the manufacturer’s instructions.

Techniques: RNA Sequencing, Derivative Assay, Activity Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Suppression of STK11 induces expansion of polymorphonuclear myeloid-derived suppressive cells and activation of immune signaling in breast cancer

doi: 10.1007/s00262-025-04189-8

Figure Lengend Snippet: Cytokine array analysis of immune-related protein expression in Stk11 -KO mouse breast cancer cell lines and their parental counterparts. A Array map. B - D Representative cytokine array blots for PY8119 B , M158 C , and NF639 D cell lines, comparing Stk11 -KO cells with their parental counterparts. E Semiquantitative analysis of protein expression with a top tenfold change. Signal intensities were measured and expressed as fold changes relative to parental cells, based on the dot intensity from the cytokine array. The Wilcoxon rank-sum test compared the difference between Stk11 -KO and parental cells. Abbreviations: CTACK/CCL27, cutaneous T cell-attracting chemokine/C–C motif chemokine ligand 27; CXCL1/KC, chemokine (C-X-C motif) ligand 1; CXCL16, chemokine (C-X-C motif) ligand 16; IL-4, interleukin-4; M-CSF, macrophage colony-stimulating factor; MCP-2/CCL2, monocyte chemoattractant protein-1/C–C motif chemokine ligand 2; MCP-5, monocyte chemoattractant protein-5; MIP-2, macrophage inflammatory protein-2; and VEGF-A, vascular endothelial growth factor-A

Article Snippet: A Mouse Cytokine Antibody Array Kit C3 (RayBiotech, Norcross, GA) was used according to the manufacturer’s instructions.

Techniques: Expressing

Journal: Clinical & Experimental Metastasis

Article Title: A CSF-1R inhibitor both prevents and treats triple-negative breast cancer brain metastases in hematogenous preclinical models

doi: 10.1007/s10585-025-10366-x

Figure Lengend Snippet: CSF-1R suppression alters microglial cytokine secretion patterns with effects on astrocytes and tumor cells. A Mouse cytokine antibody array analysis of EOC2-derived conditioned medium treated with 100 nM BLZ945 or DMSO for 72 h. The images of cytokines with marked GM-CSF, IL-1Rα, IL-6 and TNFα are shown in the left panel. The pixel density of the above cytokines relative to the internal control is shown by the graphs in the right panel. B RT-PCR analysis of GM-CSF , IL-1Rα , IL-6 , and TNFα was performed on fixed, frozen specimens from vehicle control and BLZ945 treatment settings of the 4T1-BR5 TNBC brain metastasis model mice on Days -14, 3, and 10. Values represent the fold change ± SD relative to vehicle. Each dot represents one mouse, and the line designates the group median. C 4T1-BR5 and 231-BR cells were allowed to invade through Matrigel for 12 h in the presence of control or BLZ945 treated EOC2-derived conditioned medium. Invaded cells were visualized by Hema 3 staining and counted using ImageJ software. D and E EOC2 conditioned medium promotes the growth of 4T1-BR5 and 231-BR cells under co-culture conditions. 4T1-BR5 and 231-BR cells were labelled with mCherry and EGFP, respectively. Brain trophic cancer cell numbers are quantified and calculated by flow cytometry based on the ratio between cancer cells and quantitative beads. The effect of astrocytes alone on cancer cell growth was measured in low serum medium under co-culture conditions. n ≥ 3 biologically independent experiments. *, p < 0.05 vs DMSO control; n.s., not significant by the Mann–Whitney test. F Schematic representation of microglial action in breast cancer brain metastasis. CSF-1R + microglia/macrophages regulate the secretion of cytokines that influence the behavior of astrocytes and brain metastatic (BrM) cells within the microenvironment

Article Snippet: The mouse cytokine antibody array (#ARY006, R&D Systems) was used according to the manufacturer's instructions and analyzed using the ImageJ software.

Techniques: Ab Array, Derivative Assay, Control, Reverse Transcription Polymerase Chain Reaction, Staining, Software, Co-Culture Assay, Flow Cytometry, MANN-WHITNEY