GSM8794315: CT.M.rep2; Mus musculus; OTHER
Briefly, After washing the testicular tissue three times with PBS, the tissue digested with 1 mg/mL of type IV collagenase (ThermoFisher, 17104019) at 37°C for 15 min. After the tissues settle naturally, take the supernatant for culture in D-MEM/F-12 (Gibco, C11330500BT). After 48 hours, wash away the non-adherent cells with PBS, and then conduct immunofluorescence identification for the purity of LCs. In order to study the distribution of NR2F2 in LC cells, we used Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme, TD903-01) to capture NR2F2-binding sites. The experimental process was performed according to the manufacturer's instructions. In brief, 1× 105 LC cells were prepared and immobilized on concanavalin A beads. Beads are incubated with a NR2F2 primary antibody (Abcam, ab211777, 1:50), followed by incubation with a secondary antibody anti-Rabbit IgG (Abcam, ab6702). Library was quantified by Equalbit dsDNA HS Assay Kit (Vazyme, EQ111-01) using Qubit™ 4 Fluorometer (Invitrogen, Q33226). Libraries were subjected to paired-end 150 bp sequencing on NovaSeq platform at Cosmos Wisdom.