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Series GSE114227 Query DataSets for GSE114227
Status Public on Apr 11, 2019
Title Gene expression profile of HeLa cells with stable inducible expression of DOCK10
Organism Homo sapiens
Experiment type Expression profiling by array
Summary The goal of this study was to identify changes in the gene expression profile (GEP) of HeLa cells in response to induction of expression of DOCK10 during 24 hours.
 
Overall design Cervical cancer epithelial HeLa cells, growing adherently as monolayers, were cultured in Dulbecco’s MEM supplemented with 10 % FCS, 50 U/ml penicillin, 50 U/ml streptomycin, 2,5 µg/ml amphotericin B, and 2 mM L-glutamine. At subconfluency, HeLa cells were transfected using lipofectamine with pUHD-15-1-Puro vector and, after treatment with 1 µg/ml puromycin during 2-3 weeks, a stable doxycycline (dox)-regulatable tTA-expressor clone was selected (HeLa-tTA). The HeLa-tTA clone was transfected using lipofectamine with pJAG4-HA-DOCK10.1 plasmid and, after treatment with 0.5 mg/ml G418 during 2-3 weeks, a stable dox-regulatable HA-DOCK10.1-expressor clone was selected (C33), as well as an HA-DOCK10.1-non-expressor clone (C23). The parental HeLa-tTA clone, the non-expressor HeLa-C23 clone, and the HA-DOCK10.1-expressor HeLa-C33 clone were washed free of dox and cultured for 24 h in the presence (dox+) and absence of dox (dox–). Only HeLa-C33-dox– expressed HA-DOCK10.1. Following detachment of cells with 0.05%-EDTA 0.02% in PBS, total RNAs were isolated using the miRNeasy Mini Kit (Qiagen). HeLa-C33-dox+/HeLa-C33-dox–, performed twice from biological replicates, were used as reference/test pairs, respectively, and these experiments were designated as HeLa-DOCK10. HeLa-tTA-dox+/HeLa-tTA-dox– and HeLa-C23-dox+/HeLa-C23-dox–, were used as controls for dox withdrawal, and were designated as HeLa-CTRL. References were labeled with cyanine-3-CTP, and tests with cyanine-5-CTP. The labeled cRNAs were mixed together and hybridized onto Agilent SurePrint G3 Human Gene Expression 8×60K v2 Microarrays targeting 50,599 biological features (technology 39494), using the Agilent Gene Expression Hybridization kit. After hybridization, the microarray slides were washed and scanned in an Agilent G2565CA DNA Microarray Scanner using the Agilent Scan Control software. Images were analyzed with the Agilent Feature Extraction software, which computes log ratios (test vs reference) following normalization correction by linear and Lowess methods.
 
Contributor(s) Parrado A
Citation(s) 30963125
Submission date May 09, 2018
Last update date Apr 13, 2019
Contact name Antonio Parrado
E-mail(s) antonio.parrado@carm.es
Organization name Hospital Clinico Universitario Virgen de la Arrixaca
Street address Ctra. Madrid-Cartagena, s/n
City Murcia
ZIP/Postal code 30120
Country Spain
 
Platforms (1)
GPL17077 Agilent-039494 SurePrint G3 Human GE v2 8x60K Microarray 039381 (Probe Name version)
Samples (4)
GSM3138225 HeLa_CTRL_1: HeLa-tTA clone cultured for 24 hr in the presence (dox+) and absence of dox (dox–)
GSM3138226 HeLa_CTRL_2: HeLa-C23 clone cultured for 24 hr in the presence (dox+) and absence of dox (dox–)
GSM3138227 HeLa_DOCK10_1: HeLa-C33 clone cultured for 24 hr in the presence (dox+) and absence of dox (dox–)
This SubSeries is part of SuperSeries:
GSE114243 The role of DOCK10 in the regulation of the transcriptome and aging
Relations
BioProject PRJNA470619

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE114227_HeLa_Matrix.txt.gz 752.7 Kb (ftp)(http) TXT
GSE114227_RAW.tar 23.9 Mb (http)(custom) TAR (of TXT)
Processed data included within Sample table
Processed data are available on Series record

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