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Sample GSM3990044 Query DataSets for GSM3990044
Status Public on Nov 13, 2020
Title III LUHMES GFP - MPP+
Sample type RNA
 
Source name LUHMES cell line
Organism Homo sapiens
Characteristics pd phenotype: n
Treatment protocol For differentiation of LUHMES cells towards dopaminergic neurons, cells were cultured in advanced DMEM/F12 (Life Technologies GmbH, Darmstadt, Germany) supplemented with 1% N2-Supplement, 2 mM L‑glutamine, 1 mM dibutyryl cAMP, 2 ng/mL GDNF and 1 μg/mL tetracycline. After 48 hours, cells were trypsinized and seeded with 7.5 x 104 cell/cm² in pre-coated flasks. Every 48 hours after re-seeding, fresh differentiation medium was added to the cells for six days. To induce a PD-like phenotype, LUHMES cells were treated with 10 µM 1-methyl-4-phenylpyridinium (MPP+; Sigma Aldrich, Munich, Germany) 6 days after initiation of differentiation for 48 hours. Control cells were supplemented with H2O.
Growth protocol LUHMES cells were cultured in flasks pre-coated with 50 µg/mL Poly-L-ornithin and 1 µg/mL Fibronectin. Cells were cultured in advanced DMEM/F12 (Life Technologies GmbH, Darmstadt, Germany) supplemented with 1% N2-Supplement, 2 mM L-glutamine and 40 ng/mL basic fibroblast growth factor.
Extracted molecule total RNA
Extraction protocol Total RNA including miRNAs was isolated using the Qiagen miRNeasy Kit according to manufacturers recommendations.
Label Cyanine 3
Label protocol The expression profile of 2549 human miRNAs was determined using Agilent miRNA Complete Labeling and Hyb Kit (Agilent, Cat. No 5190-0456) according to the manufacturers recommendations
 
Hybridization protocol The expression profile of 2549 human miRNAs was determined using SurePrint G3 8x60k miRNA microarrays (Agilent) according to the manufacturers recommendations
Scan protocol Arrays were scanned with the Agilent Scanner with default settings.
Description miRNA expression without MPP+ treatment of LUHMES cells
Data processing Analysis of the received fluorescence signals was done by using Agilent AGW Feature Extraction software (version 10.7.1.1, Agilent Technologies, Santa Clara, CA, USA). Normalization of background corrected values was performed using GeneSpring by biological significance analysis (version 14.9, Agilent Technologies, Santa Clara, CA, USA).
 
Submission date Jul 31, 2019
Last update date Nov 13, 2020
Contact name Lena Krammes
E-mail(s) [email protected]
Organization name Saarland University
Department Institute for Human Genetics
Street address Kirrbergerstraße
City Homburg
ZIP/Postal code 66421
Country Germany
 
Platform ID GPL24741
Series (2)
GSE135149 Validation of human microRNA target pathways enables evaluation of target prediction tools [miRNA]
GSE135151 Validation of human microRNA target pathways enables evaluation of target prediction tools

Data table header descriptions
ID_REF
VALUE normalized, log2 transformed intensity values for all samples in set

Data table
ID_REF VALUE
Blank -8.395388
NC1_00000197 -8.395388
NC1_00000215 -8.395388
NC2_00079215 -8.395388
NC2_00092197 -8.395388
NC2_00106057 -8.395388
NC2_00122731 -8.395388
NegativeControl -8.395388
dmr_285 5.8590503
dmr_3 8.366112
dmr_308 -8.395388
dmr_316 -8.395388
dmr_31a 4.3649
dmr_6 7.9965787
hsa-let-7a-3p -8.395388
hsa-let-7a-5p 3.5135875
hsa-let-7b-3p -1.8640306
hsa-let-7b-5p -0.14728117
hsa-let-7c-3p -8.395388
hsa-let-7c-5p 2.9034376

Total number of rows: 2570

Table truncated, full table size 62 Kbytes.




Supplementary file Size Download File type/resource
GSM3990044_US11153896_257015615231_S01_miRNA_107_Sep09_2_1.txt.gz 7.1 Mb (ftp)(http) TXT
Processed data included within Sample table

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