Nucleoredoxin Downregulation Reduces β-Catenin Levels and Shifts Hematopoietic Differentiation towards Myeloid Lineage In Vitro
Abstract
:1. Introduction
2. Materials and Methods
2.1. Knockdown of Nucleoredoxin
2.2. Cell Culture
2.3. Assessment of Cell Proliferation
2.4. Study of Cell Adhesion
2.5. Flow Cytometry
2.6. Western Blot
2.7. Quantitative PCR (qPCR)
2.8. Statistical Analysis
3. Results
3.1. Effect of NRX Downregulation on Megakaryocytic Differentiation of HEL Cells
3.2. The Downregulation of NRX Prompts a Decrease in β-Catenin Levels in HEL Cells
3.3. Nrx Downregulation Dramatically Decreases Murine Hematopoietic Progenitors In Vitro
3.4. Surface Markers of Mature Blood Lineages Are Altered in Nrx-Silenced Mouse Hematopoietic Progenitors In Vitro
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Target | ID | Sequence (5′-3′) |
---|---|---|
Human NRX | NRX 1 NRX 2 | AAACAGTACTTCAGTGAGA AGAAAATCATTGCCAAGTA |
Mouse Nrx | Nrx 1 Nrx 2 | AGATCATTGCCAAGTACAA ATACCGAGTCTCCAACATT |
Target Gene | Target Strand | Sequence (5′-3′) |
---|---|---|
Human NRX | Forward Reverse | ACCCAGAAGGTCTGGAGTTC CCAATGTGCGGAGAAATAGA |
Mouse Nrx | Forward Reverse | TCGTTAGTGCAGACAGGTCA TGCCTTGGATTCCATACAGT |
Human ACTB | Forward Reverse | CACCACACCTTCTACAATGA ACATGATCTGGGTCATCTTC |
Mouse Actb | Forward Reverse | CAGCCTTCCTTCTTGGGTAT TGGCATAGAGGTCTTTACGG |
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Pérez-Fernández, A.; López-Ruano, G.; Prieto-Bermejo, R.; Sánchez-Bernal, C.; Sánchez-Yagüe, J.; Hernández-Hernández, Á. Nucleoredoxin Downregulation Reduces β-Catenin Levels and Shifts Hematopoietic Differentiation towards Myeloid Lineage In Vitro. BioChem 2021, 1, 26-35. https://doi.org/10.3390/biochem1010003
Pérez-Fernández A, López-Ruano G, Prieto-Bermejo R, Sánchez-Bernal C, Sánchez-Yagüe J, Hernández-Hernández Á. Nucleoredoxin Downregulation Reduces β-Catenin Levels and Shifts Hematopoietic Differentiation towards Myeloid Lineage In Vitro. BioChem. 2021; 1(1):26-35. https://doi.org/10.3390/biochem1010003
Chicago/Turabian StylePérez-Fernández, Alejandro, Guillermo López-Ruano, Rodrigo Prieto-Bermejo, Carmen Sánchez-Bernal, Jesús Sánchez-Yagüe, and Ángel Hernández-Hernández. 2021. "Nucleoredoxin Downregulation Reduces β-Catenin Levels and Shifts Hematopoietic Differentiation towards Myeloid Lineage In Vitro" BioChem 1, no. 1: 26-35. https://doi.org/10.3390/biochem1010003