|
| Status |
Public on Aug 09, 2012 |
| Title |
siPINC1/6 (with differentiation) 1 |
| Sample type |
RNA |
| |
|
| Source name |
HC11 mammary epithelial cells
|
| Organism |
Mus musculus |
| Characteristics |
cell line: HC11
treatment: siRNA against major PINC splice forms (PINC1 and PINC6)
differentiation: differentiation-inducing agents lactogenic hormones hydrocortisone, insulin and prolactin
|
| Treatment protocol |
Lentiviral vectors (LeGO-GFP, LeGO-1.0, LeGO-1.6 or LeGO-DCR) were co-transfected with packaging vectors (pCMV-VSVG, pMDLg/pRRE, pRSV-Rev) into 293-T cells using Fugene 6 (Roche). HC11 cells were transfected with siRNAs (10 nM/well) at 60-80 % confluency in 6 well plates using Lipofectamine RNAiMAX reagent (Invitrogen). Silencer Select siRNAs (Ambion) were used to target mPINC1.0 (n273107), mPINC1.6 (n254312) and DCR2 (n254314). The controls included mock tranfection without siRNAs and a siNEG, Silencer Select Negative Control #1 (4390843). Cells were treated with or without the lactogenic hormones hydrocortisone, insulin and prolactin to induce differentiation.
|
| Growth protocol |
Cells were grown from HC11 line, a mouse mammary epithelial cell line derived from the Comma-D cell line which were isolated from a mid-pregnant Balb/c mouse
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using TRIzol (Invitrogen) and quantified using a NanoDrop (ThermoScientific).
|
| Label |
cy3
|
| Label protocol |
standard Agilent protocol
|
| |
|
| Hybridization protocol |
Hybridizations were performed using One-colour hybridization on SurePrint G3 Mouse GE 60k Microarray (Agilent)
|
| Scan protocol |
Slide were scanned in an Agilent Microarray Scanner and the data were analyzed using Agilent Feature Extraction Software (v.9.1).
|
| Description |
HC11 mammary epithelial cells, with siRNA against major PINC splice forms (PINC1 and PINC6), and with differentiation-inducing agents
|
| Data processing |
quantile normalized, background subtracted data. Agilent and Bioconductor software was used.
|
| |
|
| Submission date |
May 18, 2012 |
| Last update date |
Aug 09, 2012 |
| Contact name |
Chad Creighton |
| E-mail(s) |
creighto@bcm.tmc.edu
|
| Organization name |
Baylor College of Medicine
|
| Department |
Biostatistics, Ducan Cancer Center
|
| Street address |
One Baylor Plaza, Mail Stop: BCM305
|
| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
| |
|
| Platform ID |
GPL13912 |
| Series (1) |
| GSE38052 |
Genome-wide transcriptional effects of Pregnancy-Induced Non-coding RNA (PINC) |
|
