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Status |
Public on Apr 01, 2024 |
Title |
Fibroblasts hTERT-BJ1: Control vs Colpvp |
Sample type |
RNA |
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Channel 1 |
Source name |
Total RNA extracted from untreated fibroblasts
|
Organism |
Homo sapiens |
Characteristics |
cell line: hTERT-BJ1 cell type: Fibroblast
|
Treatment protocol |
Controls were untreated cell cultures, Colpvp were incubated for 3 h with FBS-free medium and then with 3% collagen-PVP prepared in the corresponding medium without FBS and incubated for 24 h at 37 °C and 5% CO2
|
Growth protocol |
hTERT-BJ1 were cultured with Dulbecco's modified Eagle's Medium,supplemented with 10% FBS, 2 mM L-glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
Label |
Cy3
|
Label protocol |
10 µg of total RNA for cDNA synthesis, incorporating dUTP-Alexa555 or dUTP-Alexa647 using the First-Strand cDNA labeling kit (Invitrogen). Incorporation of fluorophore was analyzed by using the absorbance at 555 or 650 nm.
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|
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Channel 2 |
Source name |
Total RNA extracted from Colpvp treated fibroblasts
|
Organism |
Homo sapiens |
Characteristics |
cell line: hTERT-BJ1 cell type: Fibroblast
|
Treatment protocol |
Controls were untreated cell cultures, Colpvp were incubated for 3 h with FBS-free medium and then with 3% collagen-PVP prepared in the corresponding medium without FBS and incubated for 24 h at 37 °C and 5% CO2
|
Growth protocol |
hTERT-BJ1 were cultured with Dulbecco's modified Eagle's Medium,supplemented with 10% FBS, 2 mM L-glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
Label |
Cy5
|
Label protocol |
10 µg of total RNA for cDNA synthesis, incorporating dUTP-Alexa555 or dUTP-Alexa647 using the First-Strand cDNA labeling kit (Invitrogen). Incorporation of fluorophore was analyzed by using the absorbance at 555 or 650 nm.
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|
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|
Hybridization protocol |
Hybridization was performed using equal amounts of labeled cDNA with the hybridization solution UniHyb (TeleChem International INC). The arrays were incubated ( 42°C, 14 hrs), and washed with 1X SCC/0.05 % SDS at RT.
|
Scan protocol |
GenePix 4100A and GenePix software were used for the acquisition and quantification of array images using 10µm resolution
|
Description |
Biological sample 1 of 2, untreated, fibroblast Biological sample 2 of 2, colpvp, fibroblast
|
Data processing |
ArrayPro Analyzer (Media Cibernetics) was used to calculate the Alexa555 and Alexa647 density mean value and background mean value for each spot. genArise (http://www.ifc.unam.mx/genarise/) was used for the microarray data performing background correction, lowess normalization, intensity filter, replicates analysis and selecting differentially expressed genes.
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Submission date |
Mar 28, 2024 |
Last update date |
Apr 01, 2024 |
Contact name |
Krötzsch Edgar |
E-mail(s) |
kroted@yahoo.com.mx
|
Organization name |
Instituto Nacional de Rehabilitación LGII
|
Lab |
Tejido Conjuntivo
|
Street address |
Calz México-Xochimilco 289, Coapa, Col. Arenal de Guadalupe, Tlalpan
|
City |
Mexico City |
ZIP/Postal code |
14389 |
Country |
Mexico |
|
|
Platform ID |
GPL34346 |
Series (1) |
GSE262736 |
Fibroblasts hTERT-BJ1: Control vs Colpvp |
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