title: MCP4_P1
organism: Mus musculus
description: Protocols: Mice were sacrificed and spleens were surgically removed and were pressed through a 70 µm cell strainer (BD Falcon) for single cell suspension preparation. Blood was obtained with cardiac puncture or from submandibular vein. Peritoneal cells were obtained by washing peritoneal cavity with 3 - 4 ml of PBS twice. Red blood cells were lysed by ACK lysing buffer and cells were then washed with PBS with 2% of fetal bovine serum (FBS). All cells were resuspended in ice cold PBS with 2% FBS for FACs staining. TC-1, tumor cells derived from mouse lung epithelial cells and transformed by human papillomavirus were cultured in vitro. Cells were washed with PBS and 4 x 105 cells were inoculated subcutaneously in the right flank with 200 µl PBS. Twenty-six days later, tumors were surgically removed Splenocytes, peripheral blood cells, peritoneal cells and tumor cells were washed with PBS and were counted. Cells were centrifuged at 400 g for 5 min and supernatant was removed. Cells pellet (< 3 x 106 cells) was resuspend in 350 µl of RLT buffer (Qiagen). RNA was extracted with RNeasy Mini kit (Qiagen) according to manufacturer's protocol. mRNA library preparation was realized following manufacturer's recommendations (KAPA mRNA HyperPrep ROCHE). Library purity/integrity were assessed using an Agilent 2200 Tapestation (Agilent Technologies, Waldbrunn, Germany).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-07-09T04:03:54Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-06-29T17:24:37Z
attributes.tag: External Id
attributes.value: SAMEA7026351
attributes.tag: INSDC center name
attributes.value: Ligue contre le Cancer
attributes.tag: INSDC first public
attributes.value: 2020-07-09T04:03:54Z
attributes.tag: INSDC last update
attributes.value: 2020-06-29T17:24:37Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-9271:MCP4_P1
attributes.tag: age
attributes.value: 8 to 10
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: clinical information
attributes.value: transfected with 4 x 105 cells TC-1 cells until tumor formation
attributes.tag: common name
attributes.value: house mouse
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: individual
attributes.value: 4-1
attributes.tag: organism part
attributes.value: peritoneum
attributes.tag: sample name
attributes.value: E-MTAB-9271:MCP4_P1
attributes.tag: scientific_name
attributes.value: Mus musculus
attributes.tag: sex
attributes.value: female
attributes.tag: strain
attributes.value: C57BL/6J