Journal: Molecular Cancer

Article Title: Androgen-regulated genes differentially modulated by the androgen receptor coactivator L-dopa decarboxylase in human prostate cancer cells

doi: 10.1186/1476-4598-6-38

Figure Lengend Snippet: Microarray analysis of DDC-regulated genes . Experimental outline of microarray studies to identify DDC downstream targets using LNCaP-DDC stable and control LNCaP-pDEST cells. In the presence of Dox ( grey triangle , + Dox) the tetracycline Tet repressor (tetR) is released from the TetO2 sequence in the promoter of the lentiviral construct containing the DDC gene. The dissociation of the tetR allows induction of transcription for the gene of interest. LNCaP-DDC and LNCaP-pDEST (vector control) cell lines were plated in medium containing 5% charcoal-stripped serum and incubated overnight. The next day, cells were treated for 48 hours under mock-induced (-Dox) and Dox-induced (+Dox) conditions, 24 h of which was in the presence or absence of 0.1 nM R1881. Total RNA samples were isolated from each condition, labeled and hybridized on the microarrays. The relative mRNA abundance of each gene was calculated as a ratio between hormone-treated (+R1881) and hormone-untreated (- R1881) samples. The comparison of the expression data obtained from LNCaP-DDC+Dox stable cells ( left ) with the expression data from the three LNCaP control cells ( right : LNCaP-DDC-Dox, LNCaP-pDEST- Dox, and LNCaP-pDEST+Dox) yielded the identification of genes that are androgen- and DDC-regulated.

Article Snippet: Microarrays of 21,521 (70-mer) human oligos representing 21,521 genes (Operon Technologies, Inc., Alameda, CA) printed on aminosilane coated microarray slides (Matrix Technologies; Hudson, NH) were supplied by the Array Facility of The Prostate Centre at Vancouver General Hospital.

Techniques: Microarray, Sequencing, Construct, Plasmid Preparation, Incubation, Isolation, Labeling, Expressing

Journal: Molecular Cancer

Article Title: Androgen-regulated genes differentially modulated by the androgen receptor coactivator L-dopa decarboxylase in human prostate cancer cells

doi: 10.1186/1476-4598-6-38

Figure Lengend Snippet: Scatter plot of entire gene set considered expressed (p values = 0.05) in the microarray analysis . The position of each dot on the scatter plot corresponds to the normalized average signal intensity (log scale) of a single gene. The normalized average signal intensity under the DDC overexpression conditions are shown on the x and y axes (controls = no DDC overexpression and DDC = DDC overexpression). The middle line indicates values that represent a DDC/controls ratio of 1.0 (similar levels of expression in both cell lines). The outer lines represent a DDC/controls ratio of 2.0 (upper line; 2-fold greater expression in DDC compared to controls) and of 0.5 (lower line; 2-fold greater expression in controls compared to DDC).

Article Snippet: Microarrays of 21,521 (70-mer) human oligos representing 21,521 genes (Operon Technologies, Inc., Alameda, CA) printed on aminosilane coated microarray slides (Matrix Technologies; Hudson, NH) were supplied by the Array Facility of The Prostate Centre at Vancouver General Hospital.

Techniques: Microarray, Over Expression, Expressing

Journal: Molecular Cancer

Article Title: Androgen-regulated genes differentially modulated by the androgen receptor coactivator L-dopa decarboxylase in human prostate cancer cells

doi: 10.1186/1476-4598-6-38

Figure Lengend Snippet: Confirmation of microarray findings by real-time RT-PCR*

Article Snippet: Microarrays of 21,521 (70-mer) human oligos representing 21,521 genes (Operon Technologies, Inc., Alameda, CA) printed on aminosilane coated microarray slides (Matrix Technologies; Hudson, NH) were supplied by the Array Facility of The Prostate Centre at Vancouver General Hospital.

Techniques: Microarray