scRNA-seq of Wild Type and Hmgcs2-/-epithelial cells

We characterized the compositions of cell populations and signaling underlying the lineage commitments of intestinal stem cells upon loss of Hmgcs2. mRNA profiles of flow sorted intestinal crypt cells from adult wild type (WT) and Hmhgcs2-/- mice were generated by 10x sequencing. Single cells were processed through the GemCode Single Cell Platform using the GemCode Gel Bead, Chip and Library Kits (10X Genomics, Pleasanton) as per the manufacturer’s protocol. In brief, single cells were sorted into 0.4% BSA–PBS. 6,000 cells were added to each channel with an average recovery rate of 1,500 cells. The cells were then partitioned into Gel Beads in Emulsion in the GemCode instrument, where cell lysis and barcoded reverse transcription of RNA occurred, followed by amplification, shearing and 5′ adaptor and sample index attachment. Libraries were sequenced on an Illumina NextSeq 500.