Overexpression of PGRMC2 in AMCs
This protocol is extracted from research article:
Reversible EMT and MET mediate amnion remodeling during pregnancy and labor
Sci Signal, Feb 11, 2020; DOI: 10.1126/scisignal.aay1486

To determine whether oxidative stress (OS)–induced functional P4 withdrawal in AMCs occurs through PGRMC2, overexpression studies were carried out. AMCs were cultured to nearly 50% confluence in DMEM/F12 medium supplemented with 10% FBS and antimicrobial agents (penicillin-streptomycin and amphotericin) and then transfected with 800 ng of green fluorescent protein (GFP)–PGRMC2 expression plasmids (table S3) with FuGENE (1:3 plasmid weight) (Promega) in Opti-MEM I Reduced Serum Medium. After 24 hours, Opti-MEM was removed, and cells were treated with control, CSE (1:50), or CSE + P4 (200 ng/ml) for 48 hours.

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