|
| Status |
Public on Feb 25, 2017 |
| Title |
SKM-1_RPL23-KD_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
SKM-1 cells transfected with vectors carrying LV-RPL23-RNAi-EGFP (KD).
|
| Organism |
Homo sapiens |
| Characteristics |
tumor stage: an acute myeloid leukaemia cell line established in the leukaemic phase during the progression from MDS to AML (MDS/AML)
|
| Treatment protocol |
lentiviral vectors carrying LV-RPL23-RNAi-EGFP (KD group) and LV-RPL23-NC-EGFP (NC group) were constructed to produce lentivirus venom (Genechem, Shanghai, China). The supernatants of different virus-producing cells were used to transfect SKM-1 cells in combination with 5 mg/ml Polybrene (Genechem, Shanghai, China) at an appropriate MOI of 20 according to the manufacturer’s instructions. GFP expression was examined by performing flow cytometry after 48 h of infection, and knockdown efficiency was testified by qRT-PCR and western blotting as over 90%.
|
| Growth protocol |
SKM-1 (an acute myeloid leukaemia cell line established in the leukaemic phase during the progression from MDS to AML (MDS/AML)) cells (Health Science Research Resources Bank, Japan) were grown in RPMI-1640 medium (Gibco, US) containing 10% heat-inactivated foetal bovine serum, 100 IU/ml penicillin, and 100 μg/ml streptomycin in a humidified atmosphere of 5% CO2 at 37℃.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Transfected SKM-1 cells were harvested 3 days after lentiviral infection. Total RNA was extracted using TRIzol® reagent (Invitrogen) and passed quality tests (NanoDrop 2000 and Agilent Bioanalyzer 2100) according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
cDNA and biotinylated cRNA were synthesized according to the manufacturer’ instructions (GeneChip 3’IVT Express Kit) from 500 ng total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, 130 ul preliminary hybrid liquid was hybridized for 16 hr at 45℃ on a GeneChip® PrimeView™ Human Gene Expression Array (Affymetrix, Santa Clara, USA). GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000.
|
| Description |
an human acute myeloid leukaemia cell line established in the leukaemic phase during the progression from MDS to AML (MDS/AML)
|
| Data processing |
Background correction, normalization, and expression calculations were performed using Robust Multichip Average (RMA).
|
| |
|
| Submission date |
Feb 24, 2017 |
| Last update date |
Feb 25, 2017 |
| Contact name |
Yuekun Qi |
| E-mail(s) |
1319678264@qq.com
|
| Organization name |
Shanghai Jiao Tong University Affiliated Sixth People’s Hospital
|
| Department |
Department of Hematology
|
| Street address |
No.600 Yishan Road
|
| City |
Shanghai |
| State/province |
Shanghai |
| ZIP/Postal code |
200233 |
| Country |
China |
| |
|
| Platform ID |
GPL15207 |
| Series (1) |
| GSE95348 |
Expression data from ribosomal protein (RP) L23 (RPL23)-KD and NC SKM-1 cells. |
|
