GSM8321949: Inflamed_H3k27ac_1; Mus musculus; OTHER
Minced prostate tissues from naïve or inflamed POET3 mice were digested with collagenase followed by trypsinization. Dissociated cells were passed through 20G needles and 40 µm cell strainers to eliminate aggregates, followed by removal of red blood cells by ACK buffer. Isolated cells were stained with fluorescence-conjugated specific antibodies for CD45, CD31, Sca-1 and CD49f. CD45-CD31-Sca-1+CD49f+ basal stem cells were sorted on theBD FACSAriaunder sterile conditions. Stem cells from 9 naïve prostates and 4 inflamed prostates were pooled for each naïve and inflamed replicate, respectively. Cleavage Under Targets and Tagmentation (CUT&Tag) was performed as per the published protocol (https://www.protocols.io/view/bench-top-cut-amp-tag-kqdg34qdpl25/v3). Barcoded i5 and i7 primers from Diagenode (24UDI for Tagmented libraries Set I) were used for the library preparation along with NEBNext Ultra II Q5 Master Mix. Libraries were amplified for 13 cycles on the thermal cycler followed by bead purication and validation using Agilent D1000 on a Agilent TAPE Station