|
| Status |
Public on Jul 06, 2021 |
| Title |
Mice treated with cisplatin Replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
RNAs that binded to the magnetic beads with anti-N6-methyadenosine (m6A) antibody
|
| Organism |
Mus musculus |
| Characteristics |
treatment: cisplatin
age: about 10 weeks
tissue: kidney
strain: 129/Sv
molecule: m6a modified RNAs
|
| Treatment protocol |
The mice in Cis-AKI group were injected intraperitoneal with a single dose of cisplatin (20 mg/kg body weight;) as described previously. Control mice received saline only.
|
| Growth protocol |
All mice were maintained in a temperature-controlled room under a 12 h light dark schedule and received food and water ad libitum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the kidney specimens using TRIzol reagent (Invitrogen, USA) in accordance with the manufacturer’s instructions.
|
| Label |
Cy5
|
| Label protocol |
Total RNAs were immunoprecipitated with anti-N6-methyadenosine(m6A) antibody. The immunoprecipitated "IP" fraction contained enriched m6A methylated RNAs, and the supernatant " Sup" fraction contained unmodified RNAs."IP" and "Sup" RNAs were amplified as cRNAs and labeled with Cy5 and Cy3 respectively using Arraystar Super RNA Labeling Kit.
|
| |
|
| Channel 2 |
| Source name |
RNAs that without binding to the magnetic beads with anti-N6-methyadenosine (m6A) antibody
|
| Organism |
Mus musculus |
| Characteristics |
treatment: cisplatin
age: about 10 weeks
tissue: kidney
strain: 129/Sv
molecule: unmodified RNAs
|
| Treatment protocol |
The mice in Cis-AKI group were injected intraperitoneal with a single dose of cisplatin (20 mg/kg body weight;) as described previously. Control mice received saline only.
|
| Growth protocol |
All mice were maintained in a temperature-controlled room under a 12 h light dark schedule and received food and water ad libitum.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the kidney specimens using TRIzol reagent (Invitrogen, USA) in accordance with the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
Total RNAs were immunoprecipitated with anti-N6-methyadenosine(m6A) antibody. The immunoprecipitated "IP" fraction contained enriched m6A methylated RNAs, and the supernatant " Sup" fraction contained unmodified RNAs."IP" and "Sup" RNAs were amplified as cRNAs and labeled with Cy5 and Cy3 respectively using Arraystar Super RNA Labeling Kit.
|
| |
|
| |
| Hybridization protocol |
Cy3 and Cy5 labeled cRNAs were combined together and hybridized to Arraystar Mouse mRNA&lncRNA Epitranscriptomic Arrays (8x60K, Arraystar) at 65°C for 17 hours in an Agilent Hybridization Oven.
|
| Scan protocol |
After washing, slides were scanned with Agilent Scanner G2505C.
|
| Description |
Biological replicate 2 of 4. Mice with kidney injury, treated with cisplatin, harvested 3 days after administration of cisplatin.
|
| Data processing |
Data was extracted using Agilent Feature Extraction software. The probe signals passing "P" or "M" QC flags in at least 4 samples were retained for further analysis.
IP (Cy5-labelled) intensities were normalized as relative intensities by the RNA spike-in calibration controls. Differentially m6A-methylated mRNAs, lncRNAs and other ncRNAs passing fold change and statistical significance cutoffs were identified and compiled.
|
| |
|
| Submission date |
Jan 19, 2021 |
| Last update date |
Jul 06, 2021 |
| Contact name |
canming li |
| E-mail(s) |
licanming1984@126.com
|
| Phone |
+8613533231640
|
| Organization name |
The Third Affiliated Hospital of Sun Yat-sen University
|
| Street address |
NO 600, Tianhe ave.
|
| City |
guangzhou |
| State/province |
guangdong |
| ZIP/Postal code |
510630 |
| Country |
China |
| |
|
| Platform ID |
GPL25915 |
| Series (1) |
| GSE165100 |
Mice kidney tissue: control vs. acute kidney injury |
|
