Indomethacin

Six different inhibitors of transduction signals involved in proliferation or apoptosis pathways were used: W7 (0.2 μM) (SIGMA); Neomycin 5 μM (SIGMA); Indomethacin (4 μg/ml) (Abcam); Verapamil 1 μM (Abcam); TMB8 10 μM (Abcam), which also inhibits the release of Ca²⁺ from intracellular stores; and bromophenacyl bromide 1 μM (Abcam). Most of the inhibitors were diluted in culture media except for bromophenacyl bromide and Indomethacin, which were diluted in DMSO to provide stock solutions which were then diluted 1:10,000 in the experimental wells, making the contribution of DMSO negligible. Parasites were preincubated for one hour with the inhibitors and then exposed to microwave treatment. After 24 h of subsequent incubation at 37°C, the parasitemia was measured by flow cytometry and the morphology of the parasites was analyzed in Giemsa smears through a light microscope.

Carbachol, methoctramine, and BRL-37344 (Sigma Aldrich), indomethacin (Abcam), denopamine (Santa Cruz), and α,β-methylene ATP (Cayman) were dissolved in DMSO, ethanol, or distilled water as appropriate. Krebs solution was composed of: (m m) 120 NaCl, 5.9 KCl, 25 NaHCO₃, 1.2 NaH₂PO₄·2H₂O, 5.5 glucose, 1.2 MgCl₂, and 2.5 CaCl₂. pH was adjusted to 7.4 by bubbling the solution with 95% O₂,−5% CO₂.