title: UN_C12
organism: Homo sapiens
description: Protocols: We used the THP-1 monocytic cell line obtained from ATCC (#TIB-202). After differentiation and stimulation, cells were harvested, loaded into Fluidigm C1 integrated fluidic circuits (IFCs) , and cell capturing in microcells was performed according to the standard recommended procedure from Fluidigm. After cell capturing, IFC wells with cells were imaged using light microscopy. THP-1 cells were cultured in RPMI 1640 medium (Biochrom, #FG1215) supplemented with 10% fetal bovine serum (FBS, Biochrom, #S0615). All treatment compounds were added to the standard growth medium. THP-1 monocytes were differentiated to macrophages with 10 nM phorbol-12-myristate-13-acetate (PMA, Sigma, #P8139-5MG) for 48 hours. Following differentiation, cells were stimulated with 100 ng/ml LPS (Sigma-Aldrich, #L5293-2ML) for 2 hours, with 250 µM palmitate (Sigma Aldrich, #P9767) for 24 hours in the presence of 10 nM PMA or left unstimulated. Reverse transcription and pre-amplification was performed without RNA extraction directly after cell lysis in C1 IFC according to the standard protocol using SMARTer Ultra Low Input kit (Clontech, #634936). Sequencing libraries were prepared using Nextera XT library preparation kit (Illumina, #FC-131-1096/#FC-131-1002) according to the standard procedure recommended by Fluidigm; 96-cell library pools, barcoded with Nextera indexes, were prepared for each treatment condition.
attributes.tag: ENA first public
attributes.value: 2017-09-28
attributes.tag: ENA last update
attributes.value: 2017-09-18
attributes.tag: External Id
attributes.value: SAMEA104305866
attributes.tag: INSDC center alias
attributes.value: Max Delbruck Center for Molecular Medicine
attributes.tag: INSDC center name
attributes.value: Max Delbruck Center for Molecular Medicine
attributes.tag: INSDC first public
attributes.value: 2017-09-28T17:01:30Z
attributes.tag: INSDC last update
attributes.value: 2017-09-18T15:27:26Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6075:UN_C12
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: THP-1
attributes.tag: cell type
attributes.value: macrophage
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: acute monocytic leukemia
attributes.tag: organism part
attributes.value: blood
attributes.tag: pre-analysis well quality
attributes.value: OK
attributes.tag: progenitor cell type
attributes.value: monocyte
attributes.tag: sample name
attributes.value: E-MTAB-6075:UN_C12