In Situ Detection of Endogenous HIV Activation by Dynamic Nuclear Polarization NMR and Flow Cytometry
Abstract
:1. Introduction
2. Results and Discussion
2.1. AMUPol is Stable in JLat T Cells
2.2. Detection of Endogenously Expressed HIV Proteins by DNP-MAS NMR
2.3. Flow Cytometric Sorting Improves Cell Homogeneity
3. Materials and Methods
3.1. Cell Lines and Cell Culture
3.2. HIV Reactivation
3.3. Sample Preparation
3.4. NMR Data Collection
3.5. EPR Analysis
3.6. Flow Cytometry Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
cRPMI | Complete Roswell Park Memorial Institute |
DNP | Dynamic nuclear polarizations |
FACS | Fluorescence-activated cell sorting |
HIV | Human immunodeficiency virus |
JLat | Jurkat latency model |
NMR | Nuclear magnetic resonance |
PKC | Protein kinase C |
TNF-α | Tumor necrosis factor alpha |
DMSO | Dimethyl sulfoxide |
GFP | Green fluorescent protein |
MAS | Magic angle spinning |
HEK | Human embryonic kidney cells |
PBS | Phosphate buffered saline |
CPMAS | Cross polarization magic angle spinning |
References
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Overall, S.A.; Price, L.E.; Albert, B.J.; Gao, C.; Alaniva, N.; Judge, P.T.; Sesti, E.L.; Wender, P.A.; Kyei, G.B.; Barnes, A.B. In Situ Detection of Endogenous HIV Activation by Dynamic Nuclear Polarization NMR and Flow Cytometry. Int. J. Mol. Sci. 2020, 21, 4649. https://doi.org/10.3390/ijms21134649
Overall SA, Price LE, Albert BJ, Gao C, Alaniva N, Judge PT, Sesti EL, Wender PA, Kyei GB, Barnes AB. In Situ Detection of Endogenous HIV Activation by Dynamic Nuclear Polarization NMR and Flow Cytometry. International Journal of Molecular Sciences. 2020; 21(13):4649. https://doi.org/10.3390/ijms21134649
Chicago/Turabian StyleOverall, Sarah A., Lauren E. Price, Brice J. Albert, Chukun Gao, Nicholas Alaniva, Patrick T. Judge, Erika L. Sesti, Paul A. Wender, George B. Kyei, and Alexander B. Barnes. 2020. "In Situ Detection of Endogenous HIV Activation by Dynamic Nuclear Polarization NMR and Flow Cytometry" International Journal of Molecular Sciences 21, no. 13: 4649. https://doi.org/10.3390/ijms21134649