|
| Status |
Public on Jul 06, 2022 |
| Title |
CD4pIC_NC08 |
| Sample type |
RNA |
| |
|
| Source name |
48h, third injection, TC-1 tumor
|
| Organism |
Mus musculus |
| Characteristics |
tissue: TC-1 tumor
gender: female
age: 8months
infection: MCMV
strain: C57Bl6
treatment: MHC-2 MCMV peptide + pI:C
injection: intratumoral
|
| Treatment protocol |
Tumor tissues were collected 48hrs after the last of three i.t. injections and flash-frozen in liquid nitrogen.
|
| Growth protocol |
injected subcutaneously 2.5x10^5 cells (TC-1)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Frozen tumors were pulverized in 1mL Trizol (Thermo Fisher) using ceramic beads in reinforced tubes and a Precellys24 Homogenizer (Bertin Instruments). A volume of 0.2mL of chloroform was added to the tumor lysate and mixed thoroughly before centrifugation at 12,000rpm for 15min at 4 ̊C. Aqueous phase was further processed using the RNeasy Mini QIAcube Kit (Qiagen). Purified RNA was analyzed and quantified using Nanodrop device (Nanodrop Products). A total of 200ng of RNA was used for gene expression analysis using the mouse nCounter PanCancer Immune Profiling Panel (Nanostring Technologies). Sample preparation and hybridization was done following manufacturer’s instruction. The gene expression data were normalized using the nSolver Analysis Software 4.0 (Nanostring Technologies) before further analysis.
|
| Label |
Null
|
| Label protocol |
Null
|
| |
|
| Hybridization protocol |
Set up a hybridization reaction at room temperature for each sample with the following components: 3 μL of Reporter CodeSet, 5 μL of hybridization buffer, up to 5 μL (25–100 ng) of sample RNA, and 2 μL of Capture ProbeSet
|
| Scan protocol |
nCounter XT CodeSet
|
| Description |
nCounter Mouse PanCancer Immune Profiling Panel
|
| Data processing |
nSolver
|
| |
|
| Submission date |
Jun 06, 2022 |
| Last update date |
Jul 06, 2022 |
| Contact name |
Nicolas Cuburu |
| Organization name |
NIH/NCI
|
| Department |
CCR
|
| Lab |
LCO
|
| Street address |
37 convent drive
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL30298 |
| Series (1) |
| GSE205574 |
Gene-expression analysis after intratumoral treatment with CMV-derived peptides |
|
