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Status |
Public on Jul 29, 2014 |
Title |
Human SZ95 sebocytes_control vs 10 μM CBD-treated (24hrs)_rep2-1 (technical repeat of rep2) |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
SZ95 sebocytes_vehicle
|
Organism |
Homo sapiens |
Characteristics |
cell line: SZ95 cell type: immortalized human sebaceous gland cells treated with: ethanol for 24hr (vehicle control)
|
Treatment protocol |
Cells were seeded to Petri-dishes. At cca. 60% confluence, cells were treated by CBD (10 μM) or by vehicle (ethanol) for 24 hrs.
|
Growth protocol |
Human immortalized SZ95 sebocytes, originated from human facial sebaceous glands (Zouboulis et al, J Invest Dermatol, 1999), were cultured in Sebomed® Basal Medium (Biochrom) supplemented with 10% fetal bovine serum (LifeTechnologies), 1 mM CaCl2, 5 ng/ml human epidermal growth factor (Sigma-Aldrich), 50 IU/ml penicillin and 50 μg/ml streptomycin (both from Teva).
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted by using Trizol following manufacturer's instructions
|
Label |
Cy3
|
Label protocol |
RNA samples (200 ng of each) were amplified and labeled using the Agilent Low Input Quick Amp Labeling Kit (Agilent Technologies) following the manufacturer’s protocol, with Spike-In controls
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Channel 2 |
Source name |
SZ95 sebocytes_CBD
|
Organism |
Homo sapiens |
Characteristics |
cell line: SZ95 cell type: immortalized human sebaceous gland cells treated with: 10 μM (-)-cannabidiol (CBD) for 24hrs
|
Treatment protocol |
Cells were seeded to Petri-dishes. At cca. 60% confluence, cells were treated by CBD (10 μM) or by vehicle (ethanol) for 24 hrs.
|
Growth protocol |
Human immortalized SZ95 sebocytes, originated from human facial sebaceous glands (Zouboulis et al, J Invest Dermatol, 1999), were cultured in Sebomed® Basal Medium (Biochrom) supplemented with 10% fetal bovine serum (LifeTechnologies), 1 mM CaCl2, 5 ng/ml human epidermal growth factor (Sigma-Aldrich), 50 IU/ml penicillin and 50 μg/ml streptomycin (both from Teva).
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted by using Trizol following manufacturer's instructions
|
Label |
Cy5
|
Label protocol |
RNA samples (200 ng of each) were amplified and labeled using the Agilent Low Input Quick Amp Labeling Kit (Agilent Technologies) following the manufacturer’s protocol, with Spike-In controls
|
|
|
|
Hybridization protocol |
The hybridization procedure was performed according to the Two-color Microarray-Based Gene Expression Analysis (Qick Amp Labeling) protocol using the Agilent Gene Expression Hybridization Kit. Briefly, 825 ng of the corresponding Cy3- and Cy5-labeled cRNA were combined and hybridized overnight (17 hours, 65 °C) to Agilent Whole Human Genome 4x44K Oligo Microarrays using Agilent’s recommended hybridization chamber and oven.
|
Scan protocol |
Fluorescence signals of Microarrays were detected by Agilent’s Microarray Scanner.
|
Description |
Second microarray of Sample set 2
|
Data processing |
To read out and process the microarray image files the Agilent Feature Extraction Software was used. R/BioConductor software (R version: 2.15.1, limma package) was used for background subtraction and LOWESS normalization. Extracted channels were quantile normalized. Weights of features were set according to Agilent's recommendations.
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Submission date |
May 12, 2014 |
Last update date |
Jul 29, 2014 |
Contact name |
Attila Oláh |
E-mail(s) |
olah.lcmp@gmail.com
|
Organization name |
University of Debrecen
|
Department |
Department of Physiology
|
Street address |
Nagyerdei krt. 98.
|
City |
Debrecen |
ZIP/Postal code |
H-4032 |
Country |
Hungary |
|
|
Platform ID |
GPL4133 |
Series (1) |
GSE57571 |
Effects of (-)-cannabidiol on human sebocytes |
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