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Up-regulation of p-SHP-2 and p-β-arrestin, but not of the corresponding housekeeping proteins, was detected in sHLA-G treated CD4+ T cells (Fig. 6, panel A).
After subtracting the background, each band was normalized to the corresponding housekeeping protein (that is, β-actin) appearing on the blot.
Mean intensities were corrected for background followed by normalization with the corresponding housekeeping gene products α-tubulin, nuclear TBP, or non-phosphorylated proteins (p38, ERK1/2) in case of phosphorylated kinases.
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Loci corresponding to housekeeping RNAs such as tmRNA, RnpB, and 4.5S RNA were among the most highly conserved, along with homologues of E. coli Spot42 [52], RyhB [53] and GcvB [54].
Transcript levels were normalized for each sample with the geometric mean of the corresponding selected housekeeping genes.
As expected from the D. melanogaster genome, we identified two Argonaute genes in D. pulex, corresponding a housekeeping endonuclease with no immune function, and an RNAi endonuclease.
Potential homologs to the corresponding Arabidopsis housekeeping genes were identified in the tomato unigen collection of the SOL Genomics Network Cornell Universityy; ) via amino acid sequence comparisons with the BLASTX application.
Two assembled sequences corresponding to housekeeping genes up-regulated following P. berghei infection, AS 313 and AS 139, share sequence identity with a cytochrome c oxidase subunit and ribosomal protein L44, respectively.
NOS expression was evaluated in each sample by dividing the optical density of the NOS band by the intensity of the optical density of the band corresponding to the housekeeping protein.
The relative abundance of target mRNA in each sample was calculated as 2 raised to the negative of its threshold cycle value times 106 after being normalized to the abundance of its corresponding 18S rRNA (housekeeping gene), (2- primer Threshold Cycle)/2- primerhreshold Cycle)x 106).
Bands were quantified using the Odyssey software (LiCor) and normalized to bands corresponding to the housekeeping Rho-GDI protein.
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