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Using cDNA microarrays hybridized to cDNAs made from thiolated mRNAs, a small set of host transcripts was identified and their expression verified by quantitative PCR and Northern and Western blot analyses.
Resistant cells were expanded and FLAG-tagged protein expression verified by western blot.
Individual clones were picked and gene expression verified by induction with 1 µg/ml doxycycline for 24 hours and anti-myc western blotting.
Cells with low to moderate GFP-AR expression were then single cell cloned and expression verified by western blot; subsequently, GFP-AR expression was found similar to or below the endogenous expression of AR in the LNCaP prostate cancer cell line [15].
* Expression verified by 454 transcriptome sequencing, **BLB = MHC Class IIB genes, *** BF = MHC Class I genes.
A miRNA microarray was performed on the three cell phenotypes and expression verified by qRT-PCR.
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The functional differences in MEC subpopulations identifiable by MMTVrtTA/H2BGFP expression were verified by gene expression analysis.
Expression of fusion protein expression was verified by western blot (Supplementary Fig. 8d, e).
The whole population of transfected cells was used, and Ras over-expression was verified by GFP expression (see "Results").
Back-complementation cell lines expressing D366N (LEDGF/p75D366N) or WT LEDGF/p75 (LEDGF/p75BC) were generated and their expression was verified by western blot (Fig. 3B).
The expression is verified by experimental results and considers the steel fiber enhancement and weakening effect.
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