All mGluR5 protein levels were normalized using the actin immunoreactivity values.
The target gene expression levels were normalized using the actin and α-tubulin genes as internal references.
The levels of total HIV-1 intracellular (unspliced and spliced) RNA were determined with a real-time PCR assay, using the expression level of the actin mRNA as the internal control.
The expression level of mRNA and miRNA was defined based on the threshold cycle (Ct), and relative expression levels were calculated using the 2-∆∆Ct method, using the expression level of β-actin mRNA and U6 small nuclear RNA as a reference gene.
In this study, the relative expression levels of five selected human nucleotide excision repair (NER) genes (ERCC1, XPB/ERCC3, XPG/ERCC5, CSB/ERCC6, and XPC) in 20 different types of human normal tissue were simultaneously measured by a new multiplex reverse transcription (RT -PCR assay using the expRT -PCR level of the betassayin gene as an internal control.
qPCR was performed using an ABI 7300 Real Time PCR System Sequence (Applied Biosystems) at 50°C for 2 min, 95°C for 10 min, followed by 40 cycles at 95°C for 15 s and 60°C for 1 min. The quantity of the target genes were normalized using the level of the β-actin gene, and expressed as relative quantities (RQ) compared with the value of the Human Genomic DNA.
The data were then normalized using the expression levels of β-actin mRNA.
Densitometry analysis of protein levels of transfected Spy1 at the time of seeding, normalized using Actin levels, show less than a 2 fold increase in the overall Spy1 protein levels with Spy1-TST.
To estimate transcript abundance, we normalized for transcript length and scaled using the expression levels of the β-actin isoform with ENSEMBL identifier ENST00000331789.
The amount of cDNA was normalized using ß-actin levels.
