|
| Status |
Public on May 27, 2019 |
| Title |
D4Ba/F3-gp130-CIR3 stimulated with GFP-PFC (exon-level) |
| Sample type |
RNA |
| |
|
| Source name |
D4Ba/F3-gp130-CIR3 stimulated with GFP-PFC
|
| Organism |
Mus musculus |
| Characteristics |
cell line: Ba/F3-gp130
cell line vendor: Ba/F3 (DSMZ no.: ACC 300)
cell type: Hyper-Interleukin-6 dependent pro-B cellline
transfection: CIR-3
stimulation: GFP-PFC (50µl/ml)
|
| Treatment protocol |
Ba/F3-gp130 cell lines were washed four times with sterile PBS and incubated in serum-free DMEM for 3 h. Equal numbers of cells (2 x 10^6) were stimulated for 1h at 37°C with 50µl GFP-PFC emulsion in 1ml serum free DMEM medium (1 h at 37°C). As a positive stimulation control Ba/F3-gp130 cells were incubated for 1h at 37°C with 100ng/ml Hyper IL-6 in serum free medium (Samples E2, E4, E6)
|
| Growth protocol |
Ba/F3 cell lines were cultured in DMEM with 1% PenStrep and 10%FCS with recombinant Hyper-IL-6 (10 ng/ml cell culture supernatant) in 10 cm cell culture plattes and were splitted in a ratio from 1:10000 to 1:5
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extraction was made with RNeasy mini kit (Qiagen, Hilden, Germany) including DNAse digestion according to the manufacturer's instructions. RNA quality was evaluated using an Agilent 2100 Bioanalyzer and only high quality RNA (RIN > 8) was used for microarray analysis.
|
| Label |
biotin
|
| Label protocol |
150 ng total RNAwas amplified using the Ambion WT Expression Kit and the WT Terminal Labeling Kit (Affymetrix, Freiburg, Germany).
|
| |
|
| Hybridization protocol |
Amplified cDNA was hybridized on Affymetrix Mouse Gene ST 1.0 arrays (Affymetrix)
|
| Scan protocol |
Detection of probe intensities was performed using a GeneChip scanner 3000 7G (GDAS 1.4 package, Affymetrix).
|
| Description |
RNA expression value derived from Expression Console software; core-exon analysis
|
| Data processing |
The data were analyzed with Affymetrix Expression Console using default analysis settings for RNA analyses for gene default and exon default .CHP files.
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| |
|
| Submission date |
Oct 26, 2017 |
| Last update date |
May 27, 2019 |
| Contact name |
Birgit Knebel |
| Organization name |
German Diabetes Center
|
| Department |
Cinical Biochemistry and Pathobiochemistry
|
| Street address |
Auf'm Hennekamp 65
|
| City |
Duesseldorf |
| ZIP/Postal code |
40225 |
| Country |
Germany |
| |
|
| Platform ID |
GPL10740 |
| Series (1) |
| GSE106215 |
Tracking of Individual Cell Populations by 19F MRI using a "Cargo Internalization Receptor (CIR)" System |
|
| Relations |
| Alternative to |
GSM2831681 (gene-level analysis) |
