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Status |
Public on Dec 08, 2021 |
Title |
PD-1 hi Iso7 |
Sample type |
RNA |
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Source name |
Mouse spleen CD8 T-cells
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Organism |
Mus musculus |
Characteristics |
strain: C56BL/6 Sex: female tissue: Mouse spleen CD8 T-cells
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Treatment protocol |
After 3 weeks of transplantation, mice were treated with aIL-10R antibody or the corresponsing isotype controls every 3 weeks by intraperitoneal injection
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Growth protocol |
C57BL/6 mice transplanted with murine CLL cells and after 5 weeks total RNA was isolated from PD-1hi effector CD8+ T-cell that were sorted by flow cytometry from spleen (Lin- TCRb+ CD8+ CD127low CD44int-hi PD-1hi)
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Extracted molecule |
total RNA |
Extraction protocol |
RNA was extracted from sorted cells using the RNeasy Micro Kit (Qiagen) according to the manufacturer’s protocol
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Label |
biotin
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Label protocol |
Biotin-labeled ss-cDNA samples for hybridization on GeneChip™ Mouse Gene 2.0 ST Arrays (Affymetrix) were prepared according to Affymetrix's recommended sample labeling procedure as described in the Gene Chip WT Pico Reagent Kit guide
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Hybridization protocol |
20 μg of cRNA was then converted to sensestrand DNA (ss-cDNA). After fragmentation and terminal labeling, 5,5 ug of biotinylated ss-cDNA were hybridized for 17 hr at 45°C on Affymetrix Mouse Gene 2.0 ST Arrays
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Scan protocol |
Microarray scanning was done using an Affymetrix GeneChip® Scanner 3000 according to GeneChip® Expression Wash, Stain and Scan Manual for Cartridge Arrays.
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Description |
Isotype replicate 2
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Data processing |
Samples were processed with Affymetrix Power Tools using 'Robust Microarray Analysis' approach for the normalization. Log2-transformed transcript-level gene expression was calculated for ‘meta’ probe sets. Quality control of the samples was carried out by examining average raw intensity signal (pm_mean) or mean absolute deviation of the residuals (all_probeset_mad_residual_mean). 'Detected above background' measure was used to filter ‘meta’ probe sets if less than 50% of probe sets were called as ‘detected’ (P value <0.05) within less than 50% of the samples in each experimental group.
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Submission date |
Aug 14, 2017 |
Last update date |
Dec 08, 2021 |
Contact name |
Martina Seiffert |
Organization name |
German Cancer Research Center
|
Department |
Department of Molecular Genetics
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Street address |
Im Neuenheimer Feld 280
|
City |
Heidelberg |
ZIP/Postal code |
69120 |
Country |
Germany |
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Platform ID |
GPL16570 |
Series (2) |
GSE102601 |
Gene expression profiling of PD-1hi effector CD8+ T-cell after IL-10R blockade |
GSE102602 |
Interleukin-10 receptor signaling promotes the maintenance of a PD-1int TCF-1+ CD8+ T cell population that sustains anti-tumor immunity |
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