title: GSM3316280: MON22: Endometrial cancer; Homo sapiens; RNA-Seq
library_construction_protocol: Monocytes Protocol:  Monocytes were isolated from healthy and cancer individuals using Fluorescent- activated cell sorting (FACS) with the following markers (CD45+/CD3-CD19-CD56- /HLA-DR+MHC II+/CD14+CD16+). Immediately after FACS sorting, all samples were centrifuged at 450 RCF for 10 min at 4°C. Cell pellets were re-suspended in 350 ul of RLT lysis buffer and RNA extracted with RNAeasy Microkit (Qiagen) according to manufacturer's instructions. Macrophages/TAMs Protocol: Isolation and Fluorescence-activated cell sorting (FACS) of human tissue macrophages was performed as described (Cassetta et al. 2016). Briefly, markers for FACS isolation of macrophages included CD45+/CD3-CD19-CD56- /CD11b+/CD14+CD163+. Immediately after sorting, all samples were centrifuged at 450 RCF for 10 min at 4C. Cell pellets were re-suspended in 350 ul of RLT lysis buffer and RNA extracted with RNAeasy Microkit (Qiagen) according to manufacturer's instructions. RNA quantity was determined by QUBIT (Invitrogen); total RNA integrity was assessed by Agilent Bioanalyzer and the RNA Integrity Number (RIN) was calculated; samples that had a RIN > 7 were selected for RNA amplification and sequencing. RNA was amplified with Ovation RNAseq Amplification kit v2 (Nugen) paired-end RNA-seq  illumina Hi-Seq
attributes.tag: GEO Accession
attributes.value: GSM3316280