Intestinal tuft cells were isolated from male C57BL/6J mice. Briefly, the proximal 5 cm of the murine small intestine was dissected, flushed, cut longitudinally, and rinsed to remove luminal contents. Segments were incubated in a 37°C shaker for 20 min in 5 ml PBS containing 2.5 mM EDTA, 0.75 mM dithiothreitol (DTT), and 10 μg/ml DNaseI. Tissues were shaken vigorously for 30s, large pieces of intestinal wall were removed, and cells were spun down at 4°C, 1200 rpm, for 5 min. Supernatant was removed and cells were re-suspended in 5 ml HBSS with 1.0 U/ml Dispase and 10μg/ml DNAse I, shaking for 10 min. Digested cells were passed through a 100 μm filter and incubated with ACK lysing buffer. Single cell suspensions were incubated on ice with mouse Fc receptor block (BD Biosciences, 1:200) followed by antigen-specific antibodies in FACS buffer. DAPI (molecular probes, 1:1000) and Annexin V (Biolegend, Pacific Blue conjugate at 1:200) were used to exclude dead and dying cells. Cells were labeled with Cd45 (Alexa Fluor 488), EpCAM (Alexa Fluor 647), and Siglec F (PE) (Biolegend, 1:200). Fluorescence Minus One (FMO) staining controls were included for gating populations of interest. Cells were FACS purified at the Salk Institute's Flow Cytometry core facility on a BD Biosciences Influx cell sorter (100-µm size nozzle, 1 x PBS sheath buffer with sheath pressure set to 20 PSI). Cells were sorted in 1-drop Single Cell sort mode for counting accuracy. Two pools of intestinal tuft cells from three individual mice were used as biological replicates. Around 10000 sorted tuft cells were crosslinked with 1% PFA in PBS at room temperature for 10 minutes, sonicated with Covaris M220 into 200-700 bp fragments, and incubated with 0.6 μg of anti-Pou2f3 antibody (Santa Cruz #SC330) or control antibody (anti-IgG Cell Signaling #2729) overnight at 4C. Protein A bead pull down (Invitrogen 10001D), washing, on-bead tagmentation (Illumina Nextera FC-121-1030), reverse crosslinking, library amplification (16-18 PCR cycles), and DNA purification were performed as previously described (Schmidl et al., 2015). The input library was prepared by direct tagmentation of genomic DNA prepared from fixed, sonicated, and reverse-cross linked mouse chromatin and then amplified and processed as the ChIP DNA. ChIPmentation method (Schmidl et al., 2015)