Flow cytometric and multimodal detection of ASC speck formation in THP-1 monocytes and murine macrophages following canonical inflammasome activation
Abstract The detection of inflammasome activation plays a critical role in understanding innate immune responses across various diseases. Apoptosis-associated speck-like protein containing a CARD (ASC) speck formation serves as a direct indicator of inflammasome assembly; however, standardized detec...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2025-08-01
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| Series: | Journal of Analytical Science and Technology |
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| Online Access: | https://doi.org/10.1186/s40543-025-00507-y |
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| author | Chaelin Lee Soonsil Hyun Inmoo Rhee |
| author_facet | Chaelin Lee Soonsil Hyun Inmoo Rhee |
| author_sort | Chaelin Lee |
| collection | DOAJ |
| description | Abstract The detection of inflammasome activation plays a critical role in understanding innate immune responses across various diseases. Apoptosis-associated speck-like protein containing a CARD (ASC) speck formation serves as a direct indicator of inflammasome assembly; however, standardized detection protocols remain scarce. In this study, we evaluated ASC speck formation in THP-1 human monocytes using flow cytometry, fluorescence microscopy, and immunoblotting under canonical NLRP3 inflammasome stimulation conditions. THP-1 cells were first primed with lipopolysaccharide (LPS) and then stimulated with nigericin (Nig), which resulted in a significant increase in ASC speck-positive monocytes (from 4.86% to 15.03%, p < 0.01). Fluorescence microscopy confirmed punctate ASC localization, and Western blot analysis revealed ASC oligomer formation. These findings demonstrate that a flow cytometry-based ASC speck assay offers a sensitive and practical method for assessing inflammasome activation, establishing a robust platform for translational immunology research. |
| format | Article |
| id | doaj-art-4bc44b5bd7f945f5a51812179e41a856 |
| institution | Kabale University |
| issn | 2093-3371 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Journal of Analytical Science and Technology |
| spelling | doaj-art-4bc44b5bd7f945f5a51812179e41a8562025-08-24T11:34:18ZengSpringerOpenJournal of Analytical Science and Technology2093-33712025-08-011611910.1186/s40543-025-00507-yFlow cytometric and multimodal detection of ASC speck formation in THP-1 monocytes and murine macrophages following canonical inflammasome activationChaelin Lee0Soonsil Hyun1Inmoo Rhee2Department of Bioscience and Biotechnology, Sejong UniversityCollege of Pharmacy, Chungbuk National UniversityDepartment of Bioscience and Biotechnology, Sejong UniversityAbstract The detection of inflammasome activation plays a critical role in understanding innate immune responses across various diseases. Apoptosis-associated speck-like protein containing a CARD (ASC) speck formation serves as a direct indicator of inflammasome assembly; however, standardized detection protocols remain scarce. In this study, we evaluated ASC speck formation in THP-1 human monocytes using flow cytometry, fluorescence microscopy, and immunoblotting under canonical NLRP3 inflammasome stimulation conditions. THP-1 cells were first primed with lipopolysaccharide (LPS) and then stimulated with nigericin (Nig), which resulted in a significant increase in ASC speck-positive monocytes (from 4.86% to 15.03%, p < 0.01). Fluorescence microscopy confirmed punctate ASC localization, and Western blot analysis revealed ASC oligomer formation. These findings demonstrate that a flow cytometry-based ASC speck assay offers a sensitive and practical method for assessing inflammasome activation, establishing a robust platform for translational immunology research.https://doi.org/10.1186/s40543-025-00507-yASC speck formationInflammasome activationFlow cytometryTHP-1 monocytesMultimodal detection |
| spellingShingle | Chaelin Lee Soonsil Hyun Inmoo Rhee Flow cytometric and multimodal detection of ASC speck formation in THP-1 monocytes and murine macrophages following canonical inflammasome activation Journal of Analytical Science and Technology ASC speck formation Inflammasome activation Flow cytometry THP-1 monocytes Multimodal detection |
| title | Flow cytometric and multimodal detection of ASC speck formation in THP-1 monocytes and murine macrophages following canonical inflammasome activation |
| title_full | Flow cytometric and multimodal detection of ASC speck formation in THP-1 monocytes and murine macrophages following canonical inflammasome activation |
| title_fullStr | Flow cytometric and multimodal detection of ASC speck formation in THP-1 monocytes and murine macrophages following canonical inflammasome activation |
| title_full_unstemmed | Flow cytometric and multimodal detection of ASC speck formation in THP-1 monocytes and murine macrophages following canonical inflammasome activation |
| title_short | Flow cytometric and multimodal detection of ASC speck formation in THP-1 monocytes and murine macrophages following canonical inflammasome activation |
| title_sort | flow cytometric and multimodal detection of asc speck formation in thp 1 monocytes and murine macrophages following canonical inflammasome activation |
| topic | ASC speck formation Inflammasome activation Flow cytometry THP-1 monocytes Multimodal detection |
| url | https://doi.org/10.1186/s40543-025-00507-y |
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