A Positive Regulatory Feedback Loop between EKLF/KLF1 and TAL1/SCL Sustaining the Erythropoiesis
Abstract
:1. Introduction
2. Result
2.1. Genome-Wide Identification of EKLF Target Genes by Microarray Hybridization and Promoter-Based ChIP-Chip Analyses
2.2. Functional and Pathway Analysis of EKLF Target Genes
2.3. Identification of Potential Transcription Factors Co-Regulating the EKLF Targets
2.4. Likelihood of Tal1 Gene as a Regulatory Target of EKLF in E14.5 Fetal Liver Cells
2.5. EKLF As an Activator of Tal1 Gene Expression during Erythroid Differentiation
2.6. Binding In Vivo of EKLF to the Upstream Promoter of Tal1 Gene
2.7. Binding In Vivo of EKLF to the Proximal CACCC Box of Tal1 Promoter-Genomic Footprinting Analysis
2.8. Requirement of the Proximal CACCC Motif for Transcriptional Activation of the Tal1 Promoter by EKLF
3. Discussion
4. Material and Methods
4.1. Generation of Eklf−/− Mice
4.2. Gene expression Profiling by Affymetrix Array Hybridization
4.3. Identification of Differentially Expressed Genes
4.4. Identification of EKLF-Bound Targets by Using NimbleGen ChIP-Chip Array Hybridization
4.5. Matching between Affymetrix Probes and NimbleGen ChIP-Chip Probes
4.6. Co-Occurrence of Binding Motifs and Relative Distance Distribution
4.7. Functional Enrichment Analysis
4.8. ChIP-qPCR
4.9. Plasmid Construction
4.10. Cell Culture, Differentiation, DNA Transfection, and Knockdown of Gene Expression
4.11. RNA Analysis
4.12. Western Blotting Analysis and Antibodies
4.13. Reporter Assay
4.14. In Vivo Genomic Footprinting
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Acknowledgments
Conflicts of Interest
References
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Hung, C.-H.; Lee, T.-L.; Huang, A.Y.-S.; Yang, K.-C.; Shyu, Y.-C.; Wen, S.-C.; Lu, M.-J.; Yuan, S.; Shen, C.-K.J. A Positive Regulatory Feedback Loop between EKLF/KLF1 and TAL1/SCL Sustaining the Erythropoiesis. Int. J. Mol. Sci. 2021, 22, 8024. https://doi.org/10.3390/ijms22158024
Hung C-H, Lee T-L, Huang AY-S, Yang K-C, Shyu Y-C, Wen S-C, Lu M-J, Yuan S, Shen C-KJ. A Positive Regulatory Feedback Loop between EKLF/KLF1 and TAL1/SCL Sustaining the Erythropoiesis. International Journal of Molecular Sciences. 2021; 22(15):8024. https://doi.org/10.3390/ijms22158024
Chicago/Turabian StyleHung, Chun-Hao, Tung-Liang Lee, Anna Yu-Szu Huang, Kang-Chung Yang, Yu-Chiau Shyu, Shau-Ching Wen, Mu-Jie Lu, Shinsheng Yuan, and Che-Kun James Shen. 2021. "A Positive Regulatory Feedback Loop between EKLF/KLF1 and TAL1/SCL Sustaining the Erythropoiesis" International Journal of Molecular Sciences 22, no. 15: 8024. https://doi.org/10.3390/ijms22158024