|
Status |
Public on Sep 01, 2007 |
Title |
CAF_2 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
NPF Control
|
Organism |
Homo sapiens |
Characteristics |
NPF Control
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was isolated from NPFs, CAFs, and NPF cyclin D1 cells using total RNA isolation kit (Qiagen).
|
Label |
Cy5
|
Label protocol |
Total RNA was amplified through one round of linear amplification using the MessageAmp aRNA kit (Ambion, Austin, TX). Sample quality and quantification was assessed by agarose gel electrophoresis and absorbance at A260. Cy3 and Cy5 labeled cDNA probes were made from 4 µg of amplified RNA.
|
|
|
Channel 2 |
Source name |
Human carcinoma associated fibroblasts CAF_2
|
Organism |
Homo sapiens |
Characteristics |
Human carcinoma associated fibroblasts CAF_2. prostate cancer status: Y; Specimen ID: CAF_2; origin: primary prostate stromal cells from prostate with carcinoma; Tissue: carcinoma associated fibroblasts;
|
Extracted molecule |
total RNA |
Extraction protocol |
RNA was isolated from NPFs, CAFs, and NPF cyclin D1 cells using total RNA isolation kit (Qiagen).
|
Label |
Cy3
|
Label protocol |
Total RNA was amplified through one round of linear amplification using the MessageAmp aRNA kit (Ambion, Austin, TX). Sample quality and quantification was assessed by agarose gel electrophoresis and absorbance at A260. Cy3 and Cy5 labeled cDNA probes were made from 4 µg of amplified RNA.
|
|
|
|
Hybridization protocol |
Two NPF cyclin D1 and two CAF samples (labeled with Cy3) were hybridized head-to-head with an NPF control sample labeled with Cy5. Probes were hybridized competitively to microarrays under a coverslip for 16 h at 63oC.
|
Scan protocol |
Fluorescent array images were collected for both Cy3 and Cy5 by using a GenePix 4000B fluorescent scanner, and image intensity data were gridded and extracted using GenePix Pro 4.1 software.
|
Description |
Human carcinoma associated fibroblasts CAF_2 from Human hybridized against NPF Control
|
Data processing |
Differences in gene expression between NPF cyclin D1/NPF and CAF/NPF groups were determined using a two-sample t-test with Significance Analysis of Microarrays (SAM) software (http://www-stat.stanford.edu/_tibs/SAM/) with a False Discovery Rate (FDR) of +/- 10% considered significant (Tusher, PNAS, 2001). Similarities in gene expression between NPF cyclin D1/NPF and CAF/NPF groups were determined using a one-sample t-test in SAM with an FDR of +/- 0.1% considered significant. These results were reduced to unique genes by eliminating all but the highest scoring clones for each gene. A Pearson correlation coefficient was calculated in Excel to assess the strength of the linear relationship between NPF cyclin D1/NPF and CAF/NPF average log2 ratios.
|
|
|
Submission date |
Feb 01, 2007 |
Last update date |
Jan 02, 2013 |
Contact name |
Denise Mauldin |
E-mail(s) |
[email protected]
|
Phone |
2066673480
|
Fax |
2066672917
|
URL |
http://www.pedb.org
|
Organization name |
Fred Hutchinson Cancer Research Center
|
Department |
Human Biology
|
Lab |
Peter Nelson
|
Street address |
1100 Fairview Ave N D4-100
|
City |
Seattle |
State/province |
WA |
ZIP/Postal code |
98109 |
Country |
USA |
|
|
Platform ID |
GPL4768 |
Series (1) |
GSE6936 |
Comparison of Normal Prostate Fibroblasts Overexpressing Cyclin D1 and Carcinoma Associated Fibroblasts |
|
Data table header descriptions |
ID_REF |
the unique identifier of the feature derived from the Array List (up to 40 characters long, contained in quotation marks). |
VALUE |
normalized log (experiment/reference) ratio data comparable across rows and samples |
X |
the X-coordinate in ?m of the center of the feature-indicator associated with the feature, where (0,0) is the top left of the image. |
Y |
the Y-coordinate in ?m of the center of the feature-indicator associated with the feature, where (0,0) is the top left of the image. |
Dia. |
the diameter in ?m of the feature-indicator. |
F635 Median |
median feature pixel intensity at wavelength #1 (635 nm). |
F635 Mean |
mean feature pixel intensity at wavelength #1 (635 nm). |
F635 SD |
the standard deviation of the feature pixel intensity at wavelength #1 (635 nm). |
B635 Median |
the median feature background intensity at wavelength #1 (635 nm). |
B635 Mean |
the mean feature background intensity at wavelength #1 (635 nm). |
B635 SD |
the standard deviation of the feature background intensity at wavelength #1 (635 nm). |
% > B635+1SD |
the percentage of feature pixels with intensities more than one standard deviation above the background pixel intensity, at wavelength #1 (635 nm). |
% > B635+2SD |
the percentage of feature pixels with intensities more than two standard deviations above the background pixel intensity, at wavelength #1 (635 nm). |
F635 % Sat. |
the percentage of feature pixels at wavelength 635 that are saturated. |
F532 Median |
median feature pixel intensity at wavelength #2 (532 nm). |
F532 Mean |
mean feature pixel intensity at wavelength #2 (532 nm). |
F532 SD |
the standard deviation of the feature intensity at wavelength #2 (532 nm). |
B532 Median |
the median feature background intensity at wavelength #2 (532 nm). |
B532 Mean |
the mean feature background intensity at wavelength #2 (532 nm). |
B532 SD |
the standard deviation of the feature background intensity at wavelength #2 (532 nm). |
% > B532+1SD |
the percentage of feature pixels with intensities more than one standard deviation above the background pixel intensity, at wavelength #2 (532 nm). |
% > B532+2SD |
the percentage of feature pixels with intensities more than two standard deviations above the background pixel intensity, at wavelength #2 (532 nm). |
F532 % Sat. |
the percentage of feature pixels at wavelength 532 that are saturated. |
Ratio of Medians (635/532) |
the ratio of the median intensities of each feature for each wavelength, with the median background subtracted. |
Ratio of Means (635/532) |
the ratio of the arithmetic mean intensities of each feature for each wavelength, with the median background subtracted. |
Median of Ratios (635/532) |
the median of pixel-by-pixel ratios of pixel intensities, with the median background subtracted. |
Mean of Ratios (635/532) |
the geometric mean of the pixel-by-pixel ratios of pixel intensities, with the median background subtracted. |
Ratios SD (635/532) |
the geometric standard deviation of the pixel intensity ratios. |
Rgn Ratio (635/532) |
the regression ratio of every pixel in a 2-feature-diameter circle around the center of the feature. |
Rgn R2 (635/532) |
the coefficient of determination for the current regression value. |
F Pixels |
the total number of feature pixels. |
B Pixels |
the total number of background pixels. |
Sum of Medians |
the sum of the median intensities for each wavelength, with the median background subtracted. |
Sum of Means |
the sum of the arithmetic mean intensities for each wavelength, with the median background subtracted. |
Log Ratio (635/532) |
log (base 2) transform of the ratio of the medians. |
F635 Median - B635 |
the median feature pixel intensity at 635 with the median background subtracted. |
F532 Median - B532 |
the median feature pixel intensity at 532 with the median background subtracted. |
F635 Mean - B635 |
the mean feature pixel intensity at 635 with the median background subtracted. |
F532 Mean - B532 |
the mean feature pixel intensity at 532 with the median background subtracted. |
F635 Total Intensity |
the sum of all pixel intensities in the feature at wavelenth 635. |
F532 Total Intensity |
the sum of all pixel intensities in the feature at wavelenth 532. |
SNR 635 |
the signal-to-noise ratio of the feature, calculated as (F635 Mean - B635 Mean)/B635 SD. |
SNR 532 |
the signal-to-noise ratio of the feature, calculated as (F532 Mean - B532 Mean)/B532 SD. |
Flags |
the type of flag associated with a feature. |
Normalize |
the normalization status of the feature (included/not included). |