Dataset features

Specifications


Application: Gene expression microarray analysis
Number of samples: 12
Release date: Jan 31 2016
Last update date: Jan 9 2018
Access: Public
Diseases: Breast Neoplasms, Neoplasms
Chemicals: Estrogens, Progesterone
Dataset link A Luminal, Steroid Receptor-Positive Breast Cancer Stem Cell that Generates Intratumoral Heterogeneity is Preserved by Progesterone and Depleted by Estradiol

Experimental Protocol


The cytokeratin 5 (CK5) and CD28 studies each contains 2 sample groups (CK5+, CK5–, CD28+, CD28-) assayed in triplicate, generating 12 individual samples (12 arrays). CK5+ and CK5- samples were generated from T47Dco cells maintained in 1nM β-Estradiol + 10nM Progesterone for 5 days. For CK5-staining, single cell suspensions were permeabilized and fixed with RNAlater (Ambion Inc.), and incubated with 2 µg anti-CK5 mAb (Novocastra; NCL-L-CK5) labeled with Zenon Alexa Fluor 488 (Invitrogen; Z-25002). Stained cells were centrifuged and resuspended in RNAse-free NST buffer containing DAPI and FACS sorted (Beckman Coulter XDP-100 MoFlo) based on CK5+ vs. CK5– expression. RNA was extracted using the PicoPure RNA isolation kit (Arcturus). RNA from T47Dco-derived CK5+ and CK5– cells were obtained in triplicate and expression profiled using Agilent 4x44K chips at MOgene LC (www.mogene.com). To isolate CD28+ and CD28- cell populations, T47Dco cells were treated with E+P for 5 days, stained for CD28 (Invitrogen; CD2801), FACS sorted and mRNA was extracted. CD28+ and CD28– gene profiles were generated as described above.

Repositories


GEO

GSE65407

BioProject

PRJNA273890

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Contact


Aaron Knox