GSM8973580: Tamoxifen 2uM, CD8, aERKLF3, rep2; Mus musculus; OTHER
EXP-1: For KLF2 CUT&Tag, P14 cells were sorted from day 8 post-infection spleens and lymph nodes. Libraries were prepared following the Bench Top CUT&Tag v3 protocol by Steven Henikoff lab (Hatice S Kaya-Okur, 2020; Kaya-Okur et al., 2019) with minor modifications. Briefly, 200,000 live cells were lysed, nuclei isolated and bound to concanavalin A beads (EpiCypher). Nuclei were incubated overnight with anti-KLF2 antibody (Millipore #09-820) or Rabbit IgG (EpiCypher #13-0042). Anti-Rabbit Secondary Antibody (EpiCypher #13-0047) and pAG-Tn5 (EpiCypher) were added the next day. Tagmentation was performed in 50 μL of Tagmentation Buffer for 1 hour at 37°C in a thermocycler. The reaction was stopped by adding 3 μL of 0.5 M EDTA, 1 μL of 10% SDS, and 1 μL of Proteinase K (25 mg/10 mL in H2O), followed by incubation at 50°C for 1 hour. DNA was extracted using Phenol:Chloroform:Isoamyl Alcohol (25:24:1, v/v) and purified with the MinElute Reaction Cleanup Kit (Qiagen). Libraries were amplified with dual indexed adaptors.