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Sample GSM1586229 Query DataSets for GSM1586229
Status Public on Jan 04, 2019
Title CD8+ Trm, lung 5
Sample type RNA
 
Source name CD8+ Trm_lung
Organism Mus musculus
Characteristics strain background: C57Bl/6
genotype/variation: wild type
gender: female
age: 8-16 weeks
tissue: Lung
cell type: CD8+ T resident memory (Trm) cells
Treatment protocol Healthy wild type C57BL/6 female mice were sacrificed to obtain the small intestine, lungs, liver and peripheral blood (as reference). Organs were disintegrated on a Gentle MACS Octo Dissociator using the Mouse Lamina Propria Dissociation, Mouse Lung Dissociation, and Mouse Tumor Dissociation kits, respectively (all from Miltenyi Biotec), according to the manufacturer’s instructions. Crude lysates were subjected to 40%/80% Percoll gradient centrifugation (Sigma Aldrich). Then, CD8b+ CD103+ Trm cells were isolated using Miltenyi’s AutoMACS Pro magnetic sorter and Multisort MACS kits, performing two subsequent rounds of positive selection. First, cells were labeled with anti-CD8b-biotin (eBioscience) and anti-CD103-PE (BD Biosciences) antibodies. Next, CD8b+ T cells were positively selected with Anti-Biotin Multisort Microbeads (Miltenyi Biotec). Microbeads were released, the cells incubated with Anti-PE Microbeads (Miltenyi Biotec), and subjected to a second round of positive selection to retrieve the CD103+ subset. As a reference, peripheral blood-derived, non-naïve CD8b+ CD62L- T cells were also isolated from the same animals by CD8b+ T cell separation and subsequent depletion of CD62L+ cells with anti-mouse CD62L microbeads (Miltenyi Biotec). As an additional control, whole organ samples were used (n=3), which were not isolated after organ disintegration.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted from MACS-separated T cells using the RNeasy Plus Micro Kit (Qiagen) following the manufacturer's recommendations. RNA yield was measured and integrity checked on an Agilent 2100 Bioanalyzer (Agilent Technologies).
Label Cy3
Label protocol Three hundred pg of total RNA, with an RNA integrity number >8.0 per sample was reverse transcribed, amplified in two rounds, and Cy3 labeled by the Arcturus RiboAmp HS PLUS, Cy3 Kit (Life Technologies) according to the manufacturer's instructions. Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
 
Hybridization protocol Amplified samples were hybridized to 4x44k mouse whole genome microarrays using the Gene Expression Hybridization Kit (Agilent)
Scan protocol Samples were scanned on an Agilent Microarray Scanner using one color scan setting for 4x44k array slides.
Description 10
Data processing The scanned images were analyzed with Feature Extraction Software 10.7 (Agilent). Raw data were quantile normalized and further analyzed by Partek GS (Partek). To identify T-cell transcripts, whole organs were also processed in parallel, and used to identify mRNAs enriched in respective T cell samples (ANOVA with FDR Correction, p<0.005, >2 fold change). To assess organ-specific differences in T cell gene expression profiles, these 6411 transcripts were subjected to multidimensional scaling (MDS), ANOVA or t-tests, as appropriate (with FDR Correction, p<0.05), to identify differentially expressed genes (DEGs).
 
Submission date Jan 16, 2015
Last update date Jan 04, 2019
Contact name Nikolett Lupsa
E-mail(s) lupsa.nikolett@gmail.com
Organization name Semmelweis University
Department Genetics, Cell-and Immunbiology
Lab MTA-SE Lendület Research Group
Street address Nagyvárad tér 4
City Budapest
State/province Pest
ZIP/Postal code 1085
Country Hungary
 
Platform ID GPL11202
Series (2)
GSE65044 Characterization of resident CD8+ Trm cell populations using whole-genome biomarker analysis
GSE65045 Comparative analysis of murine CD8+ resident memory and effector T cells in various organ environments

Data table header descriptions
ID_REF
VALUE log2 quantile normalized signal intensity

Data table
ID_REF VALUE
A_55_P1989846 8.33102
A_55_P1991598 8.32495
A_55_P2022211 9.17978
A_55_P1980764 9.53136
A_55_P1964375 12.1836
A_51_P128876 12.0725
A_55_P2121042 8.30498
A_52_P219230 8.30316
A_51_P207591 8.23052
A_55_P2131920 8.30192
A_55_P2404223 8.30217
A_55_P2101944 16.8395
A_52_P358860 9.51512
A_51_P119031 8.24113
A_51_P309854 8.25703
A_51_P343900 12.1918
A_51_P234359 8.3116
A_51_P487813 9.96049
A_52_P613977 11.9883
A_55_P1957209 8.32393

Total number of rows: 39429

Table truncated, full table size 827 Kbytes.




Supplementary file Size Download File type/resource
GSM1586229_US22502634_252665515147_S01_GE1_1100_Jul11_1_4.txt.gz 2.2 Mb (ftp)(http) TXT
Processed data included within Sample table

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