GSM9139997: Sta3_517; Mus musculus; OTHER
Ovaries were immediately dissected and homogenized for nuclear extracts using a Nuclear Extraction Kit (SN0020, Solarbio).  The CUT&Tag procedure (Vazyme, TD904-C2) began with nuclei incubation using ConA Beads Pro (10 min, RT), followed by centrifugation and supernatant removal. Nuclear-bead complexes were resuspended in 50μL chilled Antibody Buffer containing either anti-H3K4me3 (CST #9145T; 1:50) or anti-p-Stat3 (CST #9751S; 1:100) primary antibodies with gentle inversion mixing, then incubated overnight at 4°C. After secondary antibody incubation (goat anti-rabbit/mouse IgG, 1:100; 1 h, RT) and washing, tagmentation was performed using pA/G-Tnp Pro (1 h, 37°C) followed by DNA extraction, library amplification, and purification (Vazyme N411). CUT&Tag libraries were prepared using  Hyperactive Universal CUT&Tag Assay Kit (Vazyme). Libraries were amplified using 11 cycles on the thermocycler. Post amplification libraries were size selected at 280-450bp in length. Libraries were validated using the Agilent High Sensitivity DNA Kit.