Transcriptional profiling of Acute Myeloid Leukaemia (AML) cells following co-culture with anti-CD44. RNA-seq of anti-CD44 treated, paired, attached, detached and cells in monoculture using two AML cell lines, KG1a and OCI-AML3
Transcriptional profiling of Acute Myeloid Leukaemia (AML) cells following co-culture with anti-CD44. RNA-seq of anti-CD44 treated, paired, attached, detached and cells in monoculture using two AML cell lines, KG1a and OCI-AML3
OCI-AML3 and KG1a cells were pre-treated with anti-CD44 for 1 hour and then added to the co-culture for 3 hours. Cells were also treated in monoculture. Following this, the EasySep™ human CD45 Positive Selection Kit and an EasySep™ magnet were used to isolate the AML cells as per the manufacturer's instructions.  To ensure all AML cells were treated in an identical fashion, both attached, detached and AML cells in monoculture were selected. Following selection, the cells were immediately snap frozen and frozen at -80 degrees.  Total RNA was exctracted from attached (from co-culture), detached (from co-culture) and alone (from monoculture) OCI-AML3 and KG1a cells following anti-CD44 pre-treatment for 1 hour and co-culture for 3 hours. The RNA extraction was perfromed by Active Motif using the MagMAX™ mirVana™ Total RNA Isolation Kit according to the manufacturer's protocol. RNA quality was checked by RNA Screen tape and an RNA integrity number (RIN) score of >8.3 was used as a threshold. The samples were all treated with DNase. Illumina's TruSeq Stranded mRNA Library kit (Cat# 20020594). Performed by Active Motif, according to the manufacturer's protocol.