Nucleolar Stress Functions Upstream to Stimulate Expression of Autophagy Regulators
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. siRNA Sequences
2.2. Antibodies and Dyes
2.3. Cell Culture, Transfections and Drug Treatments
2.4. Cell Lysates, Western Blots and Densitometry
2.5. qRT-PCR Analysis
2.6. Immunofluorescence
2.7. Image Acquisition and GFP-LC3 Assay
2.8. Statistical Analysis
3. Results
3.1. Depletion of the Key Ribosome Biogenesis Factor PPAN Stimulates Transcription of ATG7 and ATG16L1 and Results in Increased Protein Levels of ATG7
3.2. Depletion of the Key Ribosome Biogenesis Factors NPM, SBDS or PES1 and the Transcription Factor UBF-1 Stimulates Transcription of ATG7 and ATG16L1
3.3. Depletion of the Key Ribosome Biogenesis Factors NPM, SBDS, PES1 or UBF-1 Results in Increased Protein Levels of ATG7
3.4. Interfering with RNA pol I Function by CX-5461 Stimulates Transcription of ATG7 and ATG16L1
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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si Control | si PPAN-B | |
---|---|---|
presence of gDNA | n.d. | n.d. |
PCR inhibition (CT < 30) | CT = 22.9 | CT = 23.09 |
RNA degradation |RQ2 CT|− |RQ1 CT| < 3.0 | RQ1 CT = 19.33 RQ2 CT = 20.52 =1.19 | RQ1 CT = 19.49 RQ2 CT = 20.82 =1.33 |
RT control (CT < 30) | CT = 26.51 | CT = 27.07 |
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Dannheisig, D.P.; Schimansky, A.; Donow, C.; Pfister, A.S. Nucleolar Stress Functions Upstream to Stimulate Expression of Autophagy Regulators. Cancers 2021, 13, 6220. https://doi.org/10.3390/cancers13246220
Dannheisig DP, Schimansky A, Donow C, Pfister AS. Nucleolar Stress Functions Upstream to Stimulate Expression of Autophagy Regulators. Cancers. 2021; 13(24):6220. https://doi.org/10.3390/cancers13246220
Chicago/Turabian StyleDannheisig, David P., Anna Schimansky, Cornelia Donow, and Astrid S. Pfister. 2021. "Nucleolar Stress Functions Upstream to Stimulate Expression of Autophagy Regulators" Cancers 13, no. 24: 6220. https://doi.org/10.3390/cancers13246220