1,25(OH)2D3 Promotes Macrophage Efferocytosis Partly by Upregulating ASAP2 Transcription via the VDR-Bound Enhancer Region and ASAP2 May Affect Antiviral Immunity
Abstract
:1. Introduction
2. Material and Methods
2.1. Cells Culture and In Vitro Efferocytosis Assay
2.2. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR)
2.3. Western Blot Analysis
2.4. Stable Knockdown of ASAP2
2.5. Chromatin Immunoprecipitation (ChIP) Assay
2.6. Luciferase Assay
2.7. Pulldown Assay for Testing RAC1 Activity
2.8. Immunoprecipitation Assay
2.9. Analysis of Differentially Expressed Genes (DEGs) and Functional Enrichment
2.10. The Disease Model of Experimental Autoimmune Encephalomyelitis (EAE) and Peritonitis
2.11. Fluorescence-Activated Cell Sorting (FACS) Analysis
2.12. Histological Analysis and Immunofluorescence Staining
2.13. Statistical Analyses
3. Results
3.1. ASAP2 Was Significantly Upregulated in THP-1 Cells and Macrophages following 1,25(OH)2D3 Exposure
3.2. Three Regulatory Regions in the VDR-Bound Enhancer Region of ASAP2 Regulated the Transcription of ASAP2 in Response to 1,25(OH)2D3
3.3. 1,25(OH)2D3 Promoted M2 Macrophage Efferocytosis and Transcription of Certain Genes Encoding the “Eat-me” Signals
3.4. 1,25(OH)2D3 Promoted Macrophage Efferocytosis Partly via ASAP2
3.5. The Cytoskeletal-Associated Proteins and Interactors of ASAP2
3.6. ASAP2 Promoted Interferon Signaling and Anti-Virus-Associated Pathways
3.7. Vitamin D Reduced the Number of Apoptotic Cells in EAE and Promoted Macrophage Efferocytosis in Peritonitis without Changing the Total mRNA Level of Asap2
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Shi, H.; Duan, J.; Wang, J.; Li, H.; Wu, Z.; Wang, S.; Wu, X.; Lu, M. 1,25(OH)2D3 Promotes Macrophage Efferocytosis Partly by Upregulating ASAP2 Transcription via the VDR-Bound Enhancer Region and ASAP2 May Affect Antiviral Immunity. Nutrients 2022, 14, 4935. https://doi.org/10.3390/nu14224935
Shi H, Duan J, Wang J, Li H, Wu Z, Wang S, Wu X, Lu M. 1,25(OH)2D3 Promotes Macrophage Efferocytosis Partly by Upregulating ASAP2 Transcription via the VDR-Bound Enhancer Region and ASAP2 May Affect Antiviral Immunity. Nutrients. 2022; 14(22):4935. https://doi.org/10.3390/nu14224935
Chicago/Turabian StyleShi, Hui, Jiangling Duan, Jiayu Wang, Haohao Li, Zhiheng Wu, Shuaideng Wang, Xueyan Wu, and Ming Lu. 2022. "1,25(OH)2D3 Promotes Macrophage Efferocytosis Partly by Upregulating ASAP2 Transcription via the VDR-Bound Enhancer Region and ASAP2 May Affect Antiviral Immunity" Nutrients 14, no. 22: 4935. https://doi.org/10.3390/nu14224935