Protein Expression in Human Pulmonary Artery

Proteins were extracted from human pulmonary artery tissues or PAECs and lysed in RIPA lysis buffer (RIPA, Beyotime, China) in accordance with the instructions of the manufacturer. Protein concentrations were evaluated using the BCA assay based on the albumin standard. The primary antibodies were used as follows: Dec1 and PPARγ (Abcam, United States, dilution: 1/1,000, for both), Cyclin B1 and Cyclin D1 (Sigma, United States, dilution 1/1,000, for both), Bax (Cell Signaling Technology, United States, dilution: 1/1,000), Bcl-2 (Sigma, United States, dilution 1/1,000), cleaved caspase 3, and β-actin (Santa Cruz, United States, dilution 1/1,000, for both). Detailed information of Western blot was described previously (Li et al., 2020 (link)). β-actin was used as a loading control. Fold change of protein expression was evaluated as average fold change for the ratio of targeting genes/β-actin in treated samples compared with that in the controls. Measurements were repeated three times.