human monocytic leukemia cell line  (ATCC)

Journal: iScience

Article Title: Serglycin modulates inflammation and metabolism in macrophages

doi: 10.1016/j.isci.2026.115235

Figure Lengend Snippet: SRGN deficiency disrupts macrophage inflammatory activation and alters mitochondrial function (A) Experimental design. Wild-type ( SRGN +/+ ) and knockout ( SRGN −/− ) THP-1 cells were differentiated into macrophages (M0) and polarized to M1 with LPS and IFN-γ for 24 h, followed by RNA-seq and proteomic analysis of culture supernatants ( n = 4). (B) Relative SRGN expression in wild-type THP-1 cells, showing significant induction after M1 polarization (mean ± SEM; unpaired Student’s t test; ∗∗ p < 0.0001). (C) Gene set enrichment analysis (GSEA) of the macrophage activation signature comparing SRGN −/− and SRGN +/+ THP-1 cells. Knockout cells displayed a reduced extent of activation compared to wild-type cells. (D) Differential expression analysis (volcano plot) of SRGN −/− versus SRGN +/+ THP-1 M1 cells. A total of 2,554 genes were significantly altered (adjusted p < 0.05, fold-change threshold of 1.2), with 1,112 upregulated and 1,442 downregulated in knockout cells. (E and F) Hallmark pathway enrichment analysis of significantly differentially expressed genes. Upregulated pathways in SRGN −/− THP-1 cells included inflammatory response, interferon-γ response, interferon-α response, IL2/STAT5 signaling, and epithelial-mesenchymal transition (E). Downregulated pathways included mTORC1 signaling, glycolysis, cholesterol homeostasis, bile acid metabolism, and reactive oxygen species (ROS) pathways (F). Significant pathways are marked with an asterisk (∗). (G) Overlap of differentially expressed genes between SRGN −/− THP-1 cells and Srgn −/− murine macrophages. A total of 115 commonly upregulated and 189 commonly downregulated genes were identified. (H–I) Pathway enrichment analysis of commonly altered genes. Upregulated genes were enriched for inflammatory response and interferon pathways (H), whereas downregulated genes were enriched for TNF-α/NF-κB signaling and glycolysis (I). (J) Overlap of TNF-α signaling genes between human and mouse macrophages, revealing 15 commonly upregulated and 9 commonly downregulated genes. Notably, commonly downregulated genes included ZFP36 , KLF2 , HES1 , and BHLHE40 , regulators of cytokine production and inflammatory programs.

Article Snippet: THP-1 human monocytic cell line , ATCC , RRID: CVCL_0006.

Techniques: Activation Assay, Knock-Out, RNA Sequencing, Expressing, Quantitative Proteomics

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 human monocytic cell line , ATCC , RRID: CVCL_0006.

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

Journal: iScience

Article Title: Serglycin modulates inflammation and metabolism in macrophages

doi: 10.1016/j.isci.2026.115235

Figure Lengend Snippet: SRGN knockout alters the metabolic profile of macrophages (A and B) Oxygen consumption rate (OCR; maximal respiration) in wild-type (WT) and SRGN −/− (KO) THP-1 macrophages under M0 (A) and M1 (B) conditions, with or without oleic acid (OA) supplementation. Oleic acid significantly increased mitochondrial respiration in SRGN −/− macrophages (mean ± SEM; unpaired Student’s t test; ∗∗ p < 0.001 and ∗∗∗ p < 0.0001; n = 3). (C and D) Extracellular acidification rate (ECAR) in WT (C) and SRGN −/− (D) macrophages under M0 and M1 conditions, with or without oleic acid. Oleic acid did not reverse glycolytic reprogramming in M1 macrophages; however, ECAR was altered in SRGN −/− cells (mean ± SEM; p < 0.05; n = 3). (E and F) Reactive oxygen species (ROS) levels measured by DCFDA fluorescence. ROS levels were significantly reduced in SRGN −/− macrophages compared with WT under both M0 (E) and M1 (F) polarization states (mean ± SEM; ∗ p < 0.01 and ∗∗ p < 0.001; n = 6).

Article Snippet: THP-1 human monocytic cell line , ATCC , RRID: CVCL_0006.

Techniques: Knock-Out, Fluorescence