|
| Status |
Public on Jun 14, 2023 |
| Title |
HF-ESTR 5 |
| Sample type |
RNA |
| |
|
| Source name |
cardiac left ventricle
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57Bl/6
Sex: Male
age: 3-4 months
treatment: Transverse aortic constriction + 10‐day continuous‐release pellet of 0.03 mg Estradiol/kg per day
|
| Treatment protocol |
Healthy mice with an ejection fraction of ˜60% were randomly subjected to sham or transaortic constriction (TAC) surgery as described previously. HF was achieved 6 to 8 weeks after the TAC surgery with an ejection fraction of ˜35%. Once TAC mice reached HF, mice were randomly euthanized (HF group), or were assigned to treatment of estradiol (E2) via a subcutaneous 10-day continuous-release pellet of 0.03 mg E2/kg per day (Innovative Research of America, )
|
| Extracted molecule |
total RNA |
| Extraction protocol |
For RNA extraction, hearts were powdered with a mortar and pestle on dry ice, suspended in Trizol (Invitrogen), and homogenized with a Polytron (Kinematica). Total RNA was isolated using Trizol according to manufacturer instructions. Quality control of the total RNA samples was assessed using gel electrophoresis. The samples sent to Ocean Ridge Biosciences were checked for quality by Bioanalyzer on Agilent 2100 Bioanalyzer RNA 6000 Pico Chip(s).
|
| Label |
Oyster-550 fluorescent dye
|
| Label protocol |
The total RNA was DNase digested and low-molecular weight (LMW) RNA was isolated by ultrafiltration through YM-100 columns (Millipore) and subsequent purification using the RNeasy MinElute Clean-Up Kit (Qiagen). The LMW RNA samples were 3’-end labeled with Oyster-550 fluorescent dye using the Flash Tag RNA labeling Kit (Genisphere).
|
| |
|
| Hybridization protocol |
Labeled LMW RNA samples were hybridized to the MicroRNA microarrays according to conditions recommended in the Flash Tag RNA labeling Kit manual.
|
| Scan protocol |
The microarrays were scanned on an Axon GenePix 4000B scanner, and data was extracted from images using GenePix V4.1 software.
|
| Data processing |
Raw data was background-subtracted, Log2-untransformed, and normalized. Intensity for each oligo probe is based on averaging of triplicate spots. Normalized threshold is calculated based on: log2(5* stdev of negative controls) + trim mean (log2-transformed negative control probe signals). Normalized Factor is the averge value of 20% Trim MeanNormalized TPT95 is calculated based on 95th percentile of negative control probe signal
|
| |
|
| Submission date |
Feb 10, 2023 |
| Last update date |
Jun 14, 2023 |
| Contact name |
Mansoureh Eghbali |
| E-mail(s) |
meghbali@g.ucla.edu
|
| Organization name |
UCLA David Geffen School of Medicine
|
| Street address |
10833 Le Conte Ave
|
| City |
Los Angeles |
| State/province |
CA |
| ZIP/Postal code |
90095 |
| Country |
USA |
| |
|
| Platform ID |
GPL33116 |
| Series (1) |
| GSE225067 |
Cardiac miRNA profiling in mouse heart failure that is rescued by estrogen treatment . |
|
