Ab282110

After deparaffinization and rehydration, tissuesections were treated with sodium citrate buffer (0.01 M) in a pressure cooker for 2 min to retrieve antigens. Then the sections were soaked in 3% hydrogen peroxide for 10 min to remove endogenous peroxidase. Sections were incubated with sheep serum at room temperature for 1 h to block nonspecific binding and then incubated at 4 ℃ overnight with the following primary antibodies: Rabbit anti-T-bet antibody (Proteintech, 13,700-1-AP, 1:400, USA); Mouse anti-Gata3 antibody (abcam, ab282110, 1:2000, UK); Rabbit anti-RORγt antibody (Proteintech, 13,205-1-AP, 1: 200, USA); Rabbit anti-Foxp3 antibody (Novus, NB100-39002, 1:400, USA). Following incubation with horseradish peroxidase (HRP) conjugated secondary antibody (DD13: ivision^™ Poly-HRP sheep anti-mouse/rabbit secondary antibody, TALENT, China) at room temperature for 30 min, the 3,3-diaminobenzidine tetrahydrochloride substrate chromogen solution was used to detect signals and hematoxylin was used to redye the nucleus. The images were captured and then analyzed by Image J.

PRDM14 (#83527, Cell Signaling Technology), ACTB (AC026, ABclonal), β-catenin (ab32572, Abcam), GATA3 (ab199428, Abcam), GATA3 (ab282110, Abcam), KRT7 (ab68459, Abcam), TFAP2C (ab218107, Abcam), HAND1 (D263110, Sangon Biotech), total H3 (AF0009, Beyotime), H3K27me3 (#9733, CST), SUZ12 (#3737, CST), TACSTD2-PE (363803, BioLegend), SIGLEC6-APC (FAB2859A, R&D), goat anti-mouse IgG (H + L) (115-005-003, Jackson ImmunoResearch), goat anti-rabbit IgG (H + L) (111-005-003, Jackson ImmunoResearch), goat anti-mouse IgG H&L (Alexa Fluor® 488) (ab150113, Abcam), goat anti-rabbit IgG H&L (Alexa Fluor® 647) (ab150079, Abcam), GSK126 (T2079, TargetMol), LGK974 (T2618, TargetMol), A83-01 (HY-10432A, MCE), CHIR99021 (T2310L, TargetMol), EGF(AF-100-15, Peprotech), Y27632 (T1725, TargetMol), BMP4 (120-05, Peprotech), TSA (T6270, TargetMol), and Vitamin C (Vc) (HY-B0166, MCE) were obtained.