Cathepsin X Activity Does Not Affect NK-Target Cell Synapse but Is Rather Distributed to Cytotoxic Granules
Abstract
:1. Introduction
2. Results
2.1. Regulation of Cathepsin X Expression and Activity in Activated NK-92 Cells
2.2. Cathepsin X Is Secreted upon Stimulation
2.3. Cathepsin X Localization during NK-92 Cell Degranulation
2.4. Cathepsin X Vesicular Distribution and Co-Localization with LFA-1
2.5. Cathepsin X Inhibition Does Not Perturb Formation of Stable Immunoconjugates
2.6. Cathepsin X Inhibition Does Not Perturb NK-92 Cell Lytic Activity
3. Discussion
4. Materials and Methods
4.1. Cell Lines
4.2. Cell Lysate Preparation for Measuring Cathepsin X Activity, mRNA Expression and Protein Levels
4.3. Kinetic Assay
4.4. Western Blotting
4.5. RNA Isolation and Quantitative Real-Time PCR Analysis
4.6. Collection of Concentrated Cell CULTURE Media
4.7. Imaging Flow Cytometry for Co-Localization Analysis
4.8. Immunoconjugate Stability Assay
4.9. Transient Transfection
4.10. Cytotoxicity Assay
4.11. Statistics
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
References
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Gene | Primer Pair | Sequence (5′→ 3′) |
---|---|---|
CtsZ | F R | TGAACCATGGGGCGAGAGAG AGTGCTCCTCGATGGCAAGG |
GAPDH | F R | TGCACCACCAACTGCTTAGC TGGCATGGACTGTGGTCATG |
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Jakoš, T.; Prunk, M.; Pišlar, A.; Kos, J. Cathepsin X Activity Does Not Affect NK-Target Cell Synapse but Is Rather Distributed to Cytotoxic Granules. Int. J. Mol. Sci. 2021, 22, 13495. https://doi.org/10.3390/ijms222413495
Jakoš T, Prunk M, Pišlar A, Kos J. Cathepsin X Activity Does Not Affect NK-Target Cell Synapse but Is Rather Distributed to Cytotoxic Granules. International Journal of Molecular Sciences. 2021; 22(24):13495. https://doi.org/10.3390/ijms222413495
Chicago/Turabian StyleJakoš, Tanja, Mateja Prunk, Anja Pišlar, and Janko Kos. 2021. "Cathepsin X Activity Does Not Affect NK-Target Cell Synapse but Is Rather Distributed to Cytotoxic Granules" International Journal of Molecular Sciences 22, no. 24: 13495. https://doi.org/10.3390/ijms222413495