RILP Induces Cholesterol Accumulation in Lysosomes by Inhibiting Endoplasmic Reticulum–Endolysosome Interactions
Abstract
:1. Introduction
2. Materials and Methods
2.1. Antibodies and Reagents
2.2. Expression Plasmids
2.3. Cell Culture and Transfection
2.4. Adenoviral-Mediated Gene Expression System and pSicoR/pCDH-Mediated Gene Lentivirus Expression System
2.5. GST-Pulldown and Western Blot
2.6. Transmission Electron Microscopy (TEM) and Immunoelectron Microscopy (IEM)
2.7. Immunofluorescence Microscopy
2.8. High Intelligent and Sensitive Structured Illumination Microscope (HIS-SIM)
2.9. Statistical Analysis
3. Results
3.1. RILP Interacts with ORP1L to Interfere with Its Interaction with VAP
3.2. RILP Interferes with ER–Late Endosome/Lysosome Contact
3.3. RILP Induces Cholesterol Accumulation in the Late Endosomes/Lysosomes
3.4. Accumulation of Cholesterol Inhibits ER–Endolysosomal Contact
3.5. RILP-Induced Accumulation of Cholesterol Induces Autophagy
3.6. RILP Interacts with the ORP Family to REGULATE Cholesterol Trafficking
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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Han, Y.; Liu, X.; Xu, L.; Wei, Z.; Gu, Y.; Ren, Y.; Hua, W.; Zhang, Y.; Liu, X.; Jiang, C.; et al. RILP Induces Cholesterol Accumulation in Lysosomes by Inhibiting Endoplasmic Reticulum–Endolysosome Interactions. Cells 2024, 13, 1313. https://doi.org/10.3390/cells13161313
Han Y, Liu X, Xu L, Wei Z, Gu Y, Ren Y, Hua W, Zhang Y, Liu X, Jiang C, et al. RILP Induces Cholesterol Accumulation in Lysosomes by Inhibiting Endoplasmic Reticulum–Endolysosome Interactions. Cells. 2024; 13(16):1313. https://doi.org/10.3390/cells13161313
Chicago/Turabian StyleHan, Yang, Xiaoqing Liu, Liju Xu, Ziheng Wei, Yueting Gu, Yandan Ren, Wenyi Hua, Yongtao Zhang, Xiaoxi Liu, Cong Jiang, and et al. 2024. "RILP Induces Cholesterol Accumulation in Lysosomes by Inhibiting Endoplasmic Reticulum–Endolysosome Interactions" Cells 13, no. 16: 1313. https://doi.org/10.3390/cells13161313