Cyt 173

Male mice (8 weeks old), purchased from HFK Bioscience Co., Ltd (C57BL/6J, Beijing, China), were divided into four groups after the one-week adaptation, including (i) control group, fed on a normal diet; (ii) AngII group, subject to AngII (1 μg/kg per minute, Sigma-Aldrich) for 1 week through minipumping; (iii) AngII + IL-22 group, subject to AngII and 20 μg/kg IL-22 (CYT-173, ProSpec) via peritoneal injection; and (iv) AngII + IL-22 + AG490 group, treated by AngII, IL-22, and AG490 (10 mg/kg, sc-202046, Santa Cruz, CA, USA). One week later, the animals were sacrificed after anesthesia using phenobarbital (50 mg/kg) to obtain the lung tissues.

Rat PMVECs were purchased from BeNa Culture Collection Co., Ltd. (category number BNCC338210; Peking, China). Cells were cultured in endothelial culture medium (number 1001, Sciencell) containing 5% fetal bovine serum (FBS), 1% endothelial cell growth supplement (number 1052, Sciencell) and 1% penicillin/streptomycin solution (number 0503) in 5% CO₂ at 37°C. Cells (P2–P4) cultured in ECM medium containing no FBS for 24 were divided into four groups: (i) normal control, cultured in low-serum RPMI 1640 medium containing 2% FBS; (ii) AngII group, cells cultured in low-serum RPMI 1640 medium containing 2% FBS and AngII (1 μM, Sigma-Aldrich, St. Louis, USA); (iii) AngII + IL-22 group, cells cultured in low-serum RPMI 1640 medium containing 2% FBS, AngII (1 μM, Sigma-Aldrich, St. Louis, USA), and IL-22 (20 ng/ml, CYT-173, ProSpec); and (iv) AngII + IL-22 + AG490 group, cultured in low-serum RPMI 1640 medium containing 2% FBS, AngII (1 μM, Sigma-Aldrich, St. Louis, USA), IL-22 (20 ng/ml), and AG490 (10 μM, sc-202046, Santa Cruz). After culturing for 72 hrs, the cells were subject to apoptosis analysis and determination of STAT3 expression, respectively.

Rat PMVECs purchased from Bena Culture Collection (category No. BNCC338210, Suzhou, China) were cultured using the standard method as previously described^{15 (link)}. Briefly, cells were cultured in endothelial culture medium (No. 1001, Sciencell) containing 5% fetal bovine serum (FBS, No. 0025), 1% endothelial cell growth supplement (No. 1052) and 1% penicillin/streptomycin solution (No. 0503) in 5% CO₂ at 37 °C. PMVECs (P2-4) were expanded in monolayers in cell culture bottle. The culture medium was changed to a serum-free solution for 24 h prior to usage. The cells were divided into four groups, including (i) control group; (ii) AngII group, treated by 1 μM AngII (Sigma-Aldrich, St. Louis, USA); (iii) AngII+IL-22 group, treated by 1 μM AngII and 20 ng/mL IL-22 (CYT-173, ProSpec, CA, USA); and (iv) AngII+IL-22+AG490 group, treated by AngII (1 μM)+ IL-22 (20 ng/mL) and AG490 (10 µM, sc-202046, Santa Cruz, CA, USA). The cells were incubated for 48 h before cellular apoptosis assay.