GSM8354748: Spleen germinal center B cells, K4PC; Mus musculus; OTHER
Briefly, single-cell suspensions were obatined by mincing and filtering mouse spleens and bone marrows through 70-μm cell strainers.  Cells were incubated in phosphate-buffered saline (PBS) containing 2% (w/v) FCS with indicated antibodies for 25min at 4°C. Cell sorting were performed with BD Aria II or Aria Fusion. CUT&Tag assay was conducted using Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme TD904). 1-5×103 GCB cells were collected and bound to ConA beads for 10 min at 25 °C. Cells were washed then incubated with 1 μg indicated antibodies at 4 °C overnight. Secondary antibodies were added for 1 hr at 25 °C followed by 0.04 μM pA/G–Tnp for 1 hr at 25 °C. For fragmentation, cells were incubated at 37 °C for 1hr. After stopping fragment， DNA was collected with DNA magnetic beads (Vazyme) for library construction using Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme) and TruePrep Index Kit V2 for Illumina (Vazyme).