|
| Status |
Public on Sep 20, 2022 |
| Title |
Control-2 |
| Sample type |
RNA |
| |
|
| Source name |
Heart-Control
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL6/J
Sex: Male
age: 5 months
group/treatment: Control
tissue: Heart muscle
|
| Treatment protocol |
After the induction of dyabetes with streptozotocin mice were randomly treated for the inhibitio of MAO with pargyline. See the growth protocol for more specific data.
|
| Growth protocol |
All the animal studies were performed using male C57BL6/J mice (6-7 weeks of age and at least 20 g in weight; Charles River Laboratories, Italy). T1D was induced with streptozotocin (50 mg/kg/day in citrate buffer pH 4.5) administered intraperitoneally for five consecutive days. STZ is a glucosamine-nitrosourea compound toxic to pancreatic β-cells. Mice were then randomized to receive either vehicle or MAO inhibitor pargyline (50 mg/kg/day) for 12 weeks. Blood glucose levels were measured twice a month using glucose meter (OneTouch Ultra 2) and mice with blood glucose levels ≥17 mM were considered diabetic. The following groups were examined: (i) control mice (C), (ii) STZ-treated mice (D), (iii) control mice treated with pargyline (C+P), (iv) STZ mice treated with pargyline (D+P).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from heart samples was isolated with TRIzol reagent (ThermoFisher) according to manufacturer protocol. RNA quantification was performed using the NanoDrop 1000 spectrophotometer (Nanodrop, Wilmington, DE, USA). Total RNA integrity and content of miRNAs in each sample were assessed by capillary electrophoresis using the Agilent Bioanalyzer 2100 with the RNA 6000 Nano and the Small RNA Nano LabChips, respectively (Agilent Technologies, Palo Alto, CA, USA). Only total RNA samples with an RNA integrity number > 7 and small RNAs concentration < 30% were used for miRNA or mRNA gene expression analysis.
|
| Label |
Cy3
|
| Label protocol |
200 ng of total RNA were labelled using mRNA low imput labelling and Hyb Kit (Agilent Technologies), according to the manufacturer protocol.
|
| |
|
| Hybridization protocol |
Labeled sample was dispensed onto the microarray to perform hybridization at 65°C for 17 h with 10 rpm rotation.
|
| Scan protocol |
Microarray slides were scanned using G2505C scanner (Agilent Technologies) at 3 µm resolution.
|
| Description |
Total RNA was used to label mRNA and lncRNAs using low imput labelling kit
Control-4
|
| Data processing |
Raw microarray gene expression data corresponding only to coding genes were quantile normalized and probe expression values that did not pass filter (positive and significant) were set as NA (not available). Probes with more than 50% of NA per condition were excluded from the analysis. To identify differentially expressed genes multiple t-tests were performed using adjusted Bonferroni correction and 0.05 as p-value cut off.
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| |
|
| Submission date |
Aug 05, 2022 |
| Last update date |
Sep 20, 2022 |
| Contact name |
Gerolamo Lanfranchi |
| E-mail(s) |
stefano.cagnin@unipd.it
|
| Phone |
+39-0498276219
|
| Organization name |
University of Padova
|
| Department |
CRIBI - Biotechnology Center and Biology Department
|
| Lab |
Functional Genomics Lab
|
| Street address |
Via U. Bassi, 58/B
|
| City |
Padova |
| ZIP/Postal code |
35131 |
| Country |
Italy |
| |
|
| Platform ID |
GPL21810 |
| Series (2) |
| GSE210611 |
Monoamine oxidase-dependent pro-survival signaling in diabetic hearts [total RNA] |
| GSE210612 |
Monoamine oxidase-dependent pro-survival signaling in diabetic hearts |
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