Dynamics and Endocytosis of Flot1 in Arabidopsis Require CPI1 Function
Abstract
:1. Introduction
2. Results
2.1. The Subcellular Localization of Flot1-GFP Fusion Protein in cpi1-1 Mutants
2.2. The Intensity of the Flot1-GFP Signal in cpi1-1 Mutants
2.3. The Dynamics of Flot1-GFP at the Plasma Membrane in cpi1-1 Mutants
2.4. The Endocytosis of FLOT1-GFP in cpi1-1 Mutants
3. Discussion
4. Materials and Methods
4.1. Plant Materials and Growth Conditions
4.2. Plasmid Construction and Plant Transformation
4.3. Scanning Electron Microscopy
4.4. Drug Treatments
4.5. Confocal Imaging
4.6. VA-TIRFM Live Imaging and Data Analysis
4.7. FCS
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
BAK1 | Brassinosteroid-associated kinase 1 |
BR | Brassinosteroid |
BRI1 | Brassinosteroid Insensitive 1 |
Ch | Colletotrichum higginsianum |
CPI1 | Cyclopropylsterol isomerase 1 |
DRMs | Detergent-resistant membranes |
eBL | 24-Epibrassinolide |
EIHM | Extra-invasive hyphal membrane |
EMCCD | Electron multiplying charge-coupled device |
ETI | Effector-triggered immunity |
FAA | formalin–acetic acid–alcohol |
FCS | Fluorescence cross spectroscopy |
Fen | Fenpropimorph |
flg22 | Flagelin22 |
Flot1 | Flotillin1 |
FLS2 | Flagellin sensing 2 |
HIR | Hypersensitive-induced reaction |
LSCM | Laser scanning confocal microscope |
MS | Murashige and Skoog |
MβCD | Methyl-β-cyclodextrin |
PAMPs | Pathogen-associated molecular patterns |
Phot1 | Phototropin 1 |
PI(4,5)P2 | Phosphatidylinositol 4,5-bisphosphate |
PPMP | DL-threo-1-phenyl-2-palmitoylamino-3-morpholino-1-propanol |
Pst | Pseudomonas syringae pv. tomato |
PTI | PAMP-triggered immunity |
SEM | Scanning electron microscopy |
SPFH | Stomation/prohibitin/flotillin/HIR |
SPT | Single-particle tracking |
VA-TIRFM | Variable-angle total internal reflection fluorescence microscopy |
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Cao, Y.; He, Q.; Qi, Z.; Zhang, Y.; Lu, L.; Xue, J.; Li, J.; Li, R. Dynamics and Endocytosis of Flot1 in Arabidopsis Require CPI1 Function. Int. J. Mol. Sci. 2020, 21, 1552. https://doi.org/10.3390/ijms21051552
Cao Y, He Q, Qi Z, Zhang Y, Lu L, Xue J, Li J, Li R. Dynamics and Endocytosis of Flot1 in Arabidopsis Require CPI1 Function. International Journal of Molecular Sciences. 2020; 21(5):1552. https://doi.org/10.3390/ijms21051552
Chicago/Turabian StyleCao, Yangyang, Qizouhong He, Zengxing Qi, Yan Zhang, Liang Lu, Jingyuan Xue, Junling Li, and Ruili Li. 2020. "Dynamics and Endocytosis of Flot1 in Arabidopsis Require CPI1 Function" International Journal of Molecular Sciences 21, no. 5: 1552. https://doi.org/10.3390/ijms21051552