|
| Status |
Public on May 08, 2022 |
| Title |
DS5_Vehicle-F3 |
| Sample type |
RNA |
| |
|
| Source name |
DS5_V
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
Sex: female
age: 3 months
cells: Cornea epithelia
tissue: Cornea
group: DS5_Vehicle-F3
age (months): 2-3M
|
| Treatment protocol |
Mice were exposed to desiccating stress (DS) conditions which included a drafty low humidity (<30% relative humidity) environment and received scopolamine in the drinking water (to inhibit tear secretion) . During DS, mice received twice a day eye drops of either vehicle, C1 or pan.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using the Qiagen mini plus columns according to the manufacturer's protocol. RNA quantity and quality was measured with NanoDrop and Bioanalyzer.
|
| Label |
not provided
|
| Label protocol |
not provided
|
| |
|
| Hybridization protocol |
One hundred nanograms of total RNA were hybridized with the NanoString Technologies nCounter Gene Expression GE- Mouse -Inflammation v2 Panel Codeset. Overnight hybridization occurred for 20 hours at 65°C. Removal of excess probes with magnetic bead purification was performed on the nCounter Prep Station (software v4.0.11.2) on the High Sensitivity assay. Briefly, the probe-mRNA structure was affinity purified by its 3’ end to remove excess reporter probes, then by its 5’ end to remove excess capture probes. Once unbound probes were washed away, the tripartite structure was bound to the streptavidin-coated cartridge by the biotin capture probe, aligned by an electric current (negative to positive), and immobilized. Photobleaching and fluorophore degradation was prevented with the addition of SlowFade.
|
| Scan protocol |
The cartridge containing immobilized samples was transferred to the nCounter Digital Analyzer (software v3.0.1.4) and scanned at 555 field of view (FOV). An epi-fluorescent microscope and CCD camera identified sets of fluorescent spots, which were tabulated for data output. Quality control metrics were recorded using the nSolver Analysis Software v4.0.6.2.
|
| Data processing |
Data processing was analyzed with nCounter and ROSALIND software. Data in Matrix is after normalization by ROSALIND.
|
| |
|
| Submission date |
May 06, 2022 |
| Last update date |
May 08, 2022 |
| Contact name |
Cintia S. de Paiva |
| E-mail(s) |
cintiadp@bcm.edu
|
| Phone |
7137982124
|
| Organization name |
Baylor College of Medicine
|
| Department |
Ophthalmology
|
| Lab |
de Paiva Lab
|
| Street address |
6565 Fannin Street NC505G
|
| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
| |
|
| Platform ID |
GPL20885 |
| Series (1) |
| GSE202378 |
New, Potent, Small Molecule Agonists of Tyrosine Kinase Receptors Improve Dry Eye Disease |
|
