title: PD1.hi.cells_2
organism: Homo sapiens
description: Protocols: Fibroblasts from HC skin in passage 3 were thawed and cultured in a T175 flask (Sarstedt, cat. no. 83.3911.002) in DMEM low glucose media (VWR, cat. no. 392-0407). When confluency was reached cells were trypsinized and seeded into a 96-well plate and stimulated by adding 20% supernatant from PD-1+/- cultures (n=3), IFNγ (10 µg/ml) (n=3), PD-1+/- T cells (n=3), or no stimulation (n=3). After 24 hours T cells were removed. RNA was purified from the co-cultured fibroblasts after washing with PBS and cell lysis. RNA-purification was done using the RNeasy Mini Kit (Qiagen, cat. no. 74104) (n=1 in triplicates), and the Genomic DNA was removed by using the RNase-Free DNase Set (Qiagen, cat. no. 79254).  The RNA integrity number and concentration were measured on Bioanalyzer RNA Nano chip (Agilent, cat.no. 5067-1511). The SMARTer Stranded Total RNA Sample Prep Kit - HI Mammalian (Takara, Japan, cat.no. 634876) was used for library preparation with 200ng of the purified RNA as input following the manufacturer`s protocol. The Bioanalyzer High sensitivity DNA analysis kit (Agilent, cat.no. 5067-4626) was used to determine the size of the library fragments and the Qubit dsDNA High Sensitivity Kit (Invitrogen, cat. no. Q32851) was used to quantify the library.
attributes.tag: ENA first public
attributes.value: 2025-11-27
attributes.tag: INSDC center name
attributes.value: Omiics, Aarhus, Denmark
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-15969:PD1.hi.cells_2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: fibroblast
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: human
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: sample name
attributes.value: E-MTAB-15969:PD1.hi.cells_2
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: stimulus
attributes.value: co-culture with PD-1 hi T cell