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It plays a role in programmed cell death and possesses important housekeeping and quality control functions that contribute to health and longevity (for review, see [50] [53].
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Basal autophagy primarily functions as a housekeeping mechanism, ensuring proper turnover and quality control of macromolecules and organelles.
Additionally, control genes are included in each array to control for genomic DNA contamination, RNA quality, housekeeping and general PCR performance.
Expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a housekeeping gene, was analyzed for normalization purposes and quality controls.
Cyclophilin (CYC) and 18S ribosomal RNA (18S) were included as housekeeping genes to serve as internal controls for quantity and quality of cDNA going into the RT reaction.
Previous studies have demonstrated that AMP‐activated protein kinase (AMPK) controls autophagy through the mammalian target of rapamycin (mTOR) and Unc‐51 like kinase 1 (ULK1/Atg1) signaling, which augments the quality of cellular housekeeping, and that β‐guanidinopropionic acid (β‐ GPA), a creatine analog, leads to a chronic activation of AMPK.
RNA was derived from microdissected tumours and quality tested by confirming expression of at least three housekeeping genes for each sample as previously described (Rogerson et al, 2008).
Values from four housekeeping genes (Tbp, Hprt1, Actb and 18 sRNA) were used for loading normalization and quality screening for each sample.
Presence and quality of DNA was assured by amplification of the beta-globin housekeeping gene.
All microarray scans were inspected visually and checked for quality on the basis of the performance of negative, housekeeping and externally added Cy3/Cy5-prelabeled spiking controls.
5'/3' ratio of housekeeping genes and RNA quality vary from 0 to 1 with 0 stands for lower bound of RNA quality and 1 stands for upper bound of RNA quality.
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