Expression profiling by high throughput sequencing
The goal of the study was to sequence mRNA from tuft cells (identified as CD11c-;CD45-;EpCAM+;IL-25+ using Flare25 reporter mice) in C57BL/6 and Aire-/- mice. The data were used to investigate thymic tuft cell heterogeneity with and without AIRE protein expression.
Single cells prepared from pooled, digested, thymi were stained for flow cytometry and individual cells were sorted into wells of a 96 well plate containg 5 ul of lysis buffer. As controls, several wells were left blank (0 cells) or 100 cells were sorted.