Anti facl4

Proteins were extracted from frozen placental tissues and HTR8/ SVneo cells with RIPA lysis buffer (Beyotime, Shanghai, China). Following the manufacturer's protocol, the protein concentration was determined using a BCA Protein Assay kit (Beyotime). Western blot analysis was performed based on the technique established in our laboratory [26] . As previously described, protein samples were subjected to SDS polyacrylamide gel electrophoresis, resolved by electrophoresis, and transferred to polyvinylidene difluoride membranes (Millipore, Billerica, USA) [27] (link). The antibodies used were anti-FAM134B (1:1000 dilution; Abcam), anti-IP3R (1:1000 dilution; Abcam), anti-FACL4 (1:1000 dilution; Abcam), anti-cytochrome C (1:500 dilution; Abcam), anti-cleaved caspase-3 (1:1000 dilution; Abcam), anti-IP3R (1:1000 dilution; Abcam), anti-calnexin (1:1000 dilution; Abcam), anti-BNIP3 (1:1000 dilution; Abcam), anti-PSME2 (1:1000 dilution; Abcam), and anti-β-actin (1:1000 dilution; Abcam). All incubation was performed at 4°C overnight. The polyvinylidene difluoride membranes were then incubated with a secondary antibody conjugated with horseradish peroxidase (1:5000 dilution; Abcam). The protein bands were visualized using western bright ECL kit (Advansta, Menlo Park, USA) and quantified with the ChemiDoc™ XRS+(Bio-Rad, Hercules, USA).