|
| Status |
Public on Mar 13, 2018 |
| Title |
SAMPLE 63: male_FA_3h_rep5 |
| Sample type |
RNA |
| |
|
| Source name |
male mouse exposed to filtered air
|
| Organism |
Mus musculus |
| Characteristics |
tissue: lung tissue homogenate
exposure time: 3 hours
Sex: male
estrous cycle stage: n/a
|
| Treatment protocol |
Male and female C57BL/6 mice of 8 weeks of age were exposed to 2ppm ozone (O3) or filtered air (FA) for 3 hours. Females were exposed at different stages of the estrous cycle: diestrus 1 (D1), diestrus 2 (D2), proestrus (P), estrus (E). Total RNA was extracted from whole lung homogenates and PCR arrays were used to determine the expression of 84 miRNAs associated with inflammation and immunity (3-9 animals per condition).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Lung tissue was homogenized in Trizol. Total RNA was extracted with the Direct-zol (Zymo Research)
|
| Label |
SYBR Green
|
| Label protocol |
Small RNAs were retro-transcribed from 200 ng of total RNA using the miScript II RT kit (Qiagen). The expression of 84 mouse miRNAs predicted to regulate inflammatory genes was assayed with the Mouse Inflammatory Response & Autoimmunity miRNA PCR Array (MIMM-105Z, Qiagen)
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Data processing |
Data were analyzed in excel using Ct values for each sample, normalized to he average Ct of five miRNA housekeeping controls (SNORD61, SNORD68, SNORD72, SNORD95, SNORD96A, RNU6-2)
Matrix normalized template shows target miRNA Ct values normalized to housekeeping miRNAs as ΔCt = (Ct_Target − Ct_housekeeping)
For fold change calculations, ΔΔCt based fold-change values were obtained using sample 27 as control (2^-ΔΔCt, where -ΔΔCt = -[ΔCttest -ΔCtcontrol])). A fold change of 200 was considered as cutoff.
|
| |
|
| Submission date |
Mar 12, 2018 |
| Last update date |
Mar 13, 2018 |
| Contact name |
Patricia Silveyra |
| E-mail(s) |
psilveyr@iu.edu, ptrcslvr@gmail.com
|
| Organization name |
Indiana University Bloomington
|
| Department |
Environmental and Occupational Health
|
| Lab |
Silveyra lab
|
| Street address |
2819 E 10th St
|
| City |
Bloomington |
| State/province |
Indiana |
| ZIP/Postal code |
47408 |
| Country |
USA |
| |
|
| Platform ID |
GPL24720 |
| Series (1) |
|
