|
Status |
Public on Mar 30, 2024 |
Title |
PATIENT_#17'S_MOTHER |
Sample type |
RNA |
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|
Source name |
Stimulated T cells
|
Organism |
Homo sapiens |
Characteristics |
gender: female genotype: PSMC3 +/+ (wild-type) phenotype: Clinically healthy
|
Treatment protocol |
PBMC were stimulated with 1 µg/µl L-PHA (Sigma) and 150 U/ml IL-2 (Miltenyi Biotec) for three weeks to expand naive T cells following the procedure of Fonteneau et al.
|
Growth protocol |
PBMC were cultured in U-bottom 96-well plates together with irradiated feeder cells using RPMI 1640 medium supplemented with 10% human AB serum.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated from resting T cells using the kit from Analytic Jena AG following the manufacturer’s instructions.
|
Label |
NanoString CodSets
|
Label protocol |
Labelling was performed using gene-specific biotin-tagged molecular barcodes (CodeSets) designed and provided by the manufacturer.
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|
|
Hybridization protocol |
One hundred nanograms total RNA was hybridized with the nCounter Autoimmune Profiling panel CodeSets for 16 hours at 65°C.
|
Scan protocol |
Hybridized probes were immobilized on streptavidin-coated cartridges for digital counting by a nCounter™ Digital Analyzer.
|
Data processing |
RCC files were analysed using nSolver 4.0. Normalization of RNA loading was performed using the geometric mean of a unique housekeeping gene.
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|
|
Submission date |
Mar 31, 2023 |
Last update date |
Mar 30, 2024 |
Contact name |
Frédéric Ebstein |
E-mail(s) |
frederic.ebstein@gmail.com
|
Organization name |
French Institute of Health and Medical Research (Inserm)
|
Department |
L'unité de recherche de l'institut du thorax, Inserm UMR 1087
|
Street address |
IRS-UN - 8 quai Moncousu - BP 70721
|
City |
Nantes |
ZIP/Postal code |
44007 |
Country |
France |
|
|
Platform ID |
GPL33300 |
Series (1) |
GSE228675 |
Transcriptional analysis of T cells derived from individuals with neurodevelopmental disorder carrying de novo heterozygous PSMC3 variants. |
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