Progesterone, Mifepristone, and rTs-MAPRC2-Ab Assay

The selected concentration of progesterone (P30 ng/mL), mifepristone (M100 ng/mL), and rTs-MAPRC2-Ab (rat antisera against rTs-MAPRC2) ratio (1:300 dilution) were used at F-AL and ML stages with both controls (only RPMI and EtOH-RPMI), collected as described above, and the previously described procedures were followed according to [23 (link),24 (link)]. The 2000 worms/well of both stages (F-AL, ML) were cultured in a 6-well plate, and a 6-well precondition was used. The cultured medium consists of RPMI-1640, 10% heat-inactivated FBS (Fetal bovine serum), and 2% antibiotics (100 U/mL penicillin; 100 mg/mL streptomycin) (Gibco, Paisley, UK), and incubated at 37 °C and 5% CO₂ for 48 h and the medium changed after 24 h for all treatments. The female (F-AL) and male (M-AL) adult worms were separated under light microscopy using Axiovert Zeiss Microscope (25× Neo Plan objectives) and observed the phenotypic appearance at both stages (F-AL and ML) using an inverted bright field microscope (Olympus, Shibuya, Japan). In the F-AL stage, the pregnancy maintenance or aborted to NBL were observed. While at the ML stage ecdysis (molting process) and motility of parasites were noted.