GSM8263915: dLAM1-CSF; Mus musculus; RNA-Seq
Fluorescence-activated cell sorting (FACS) was used to prepare cell samples for bulk RNA-seq. Single cell suspensions from CSF of Cx3cr1CreER:mTmG (tamoxifen treat) LM models were prepared and stained. Cells were sorted using the MA900 Cell Sorter into DMEM medium with 10% FBS and antibiotics. CD45+CD11b+F4/80+Ly6C-GFP+ cells were sorted as dLAMs. CD45+CD11b+F4/80+GFP- and CD45+CD11b+F4/80lowLy6C+GFP- cells were sorted and mixed together as monocyte-macrophages. Total RNA from dLAMs or Mono-Macrophage was isolated with RNeasy Mini Kit (Cat#74106, Qiagen). RNA quality and quantity was assessed by Bioanalyzer 4200 (Agilent technologies). All samples with RNA integrity (RIN) value >7 were prepared for library construction and RNA-seq on a NovaSeq 6000 platform (Illumina) using paired-end 150-bp reads, to a sequencing depth of >10 million reads per sample.