GSM5292444: ChAT-eGFPneg_IWAT_macrophages_2; Mus musculus; RNA-Seq
IWAT was harvested and digested to yield the stromal vascular fraction (SVF). SVF cells were stained with fluorescently-conjugated antibodies and ChAT-eGFP+ or ChAT-eGFP- macrophages (live CD45+ CD11b+ CD64+ Ly6G- SiglecF- NK1.1- CD3- CD19-) were isolated by FACS. Cells were sorted directly into NEBNext Lysis Buffer containing RNase inhibitor then flash frozen. Library preparation was undertaken using the NEBNext Single Cell/Low Input RNA Library Prep Kit (NEB #E6420) according to manufacturer's instructions. Quality control assessment on prepared libraries was performed using Agilent TapeStation. Paired end (150 bp) sequencing was performed on an Illumina NovaSeq (S4).