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Status |
Public on Sep 04, 2022 |
Title |
Control, PBMCs treated with HFn-BDC 3 |
Sample type |
SRA |
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Source name |
F67, HFn, BDC
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Organism |
Homo sapiens |
Characteristics |
individual: F67 disease state: Control tissue: Blood cell type: PBMC treatment: HFn-BDC
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Treatment protocol |
PBMCs were independently treated for 48 h with Hfn-BDC (10 μM). The experiment is composed by 4 different condition: AD not treated (AD NT), Controls not treated (CTR NT), AD treated with HfnBDC (AD HfnBDC) and controls treated with HfnBDC (CTR HfnBDC). After treatment, cell viability was assayed by a trypan blue exclusion test.
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Growth protocol |
Peripheral blood mononuclear cells (PBMCs) were prepared by centrifugation. Peripheral blood was layered (density=1.077) and centrifuged at 950g for 30 min. After isolation on a Ficoll-Histopaque layer (Sigma, Italy), cell viability was assayed by a trypan blue exclusion test and the cells were used for RNA extraction.
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Extracted molecule |
total RNA |
Extraction protocol |
Samples were homogenized and total RNA was isolated by Trizol® reagent(Life Science Technologies, Italy) following the manufacturer’s specifications. RNAs were quntified using a Nanodrop ND-100 Spectrophotometer (Nanodrop Technologies, Wilmington, USA) and a 2100 Bioanalyzer (Agilent RNA 6000 Nano Kit, Waldbronn, Germany); RNAs with a 260:280 ratio of ≥1.5 and an RNA ntegrity number of ≥8 were subjected to deep sequencing. Sequencing libraries were prepared by the Illumina TruSeq Stranded Total RNA kit (Illumina) using 500-ng total RNA (Illumina) and by the Lexogen SENSE Total RNA-Seq Library Prep Kit using 500-ng total RNA (Lexogen). Qualities of sequencing libraries were assessed with 4200 TapeStation with the DNA1000 reagnt kit. RNA processing has been carried out using Illumina NextSeq 500 Sequencing.
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Library strategy |
RNA-Seq |
Library source |
transcriptomic |
Library selection |
cDNA |
Instrument model |
Illumina NextSeq 500 |
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Data processing |
FastQ files were generated via llumina bcl2fastq2 Version 2.17.1.14 Gene and transcript intensities were computed using STAR/RSEM software using Gencode Release m24 (GRCm38) as a reference, using the “-strandness forward” option. Differental expression analysis for mRNA and lncRNA was performed using R package DESeq2 Assembly: HG19 Supplementary files format and content: counts, FPKMs, TPMs
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Submission date |
May 19, 2022 |
Last update date |
Sep 04, 2022 |
Contact name |
Maria Garofalo |
E-mail(s) |
maria.garofalo01@universitadipavia.it
|
Organization name |
IRCCS Mondino Foundation
|
Lab |
Genomic and Post Genomic Unit
|
Street address |
Via Mondino 2
|
City |
Pavia |
ZIP/Postal code |
27100 |
Country |
Italy |
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Platform ID |
GPL18573 |
Series (1) |
GSE203408 |
BisDemethoxyCurcumin (BDC) loaded H-Ferritin-Nanocages Mediate Regulation of inflammation in Alzheimer’s Disease Patients |
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Relations |
BioSample |
SAMN28554652 |
SRA |
SRX15387293 |