1 3 bis

Parasite suspensions (4 ×10⁶ promastigotes/mL) were treated with 4.1, 15.0, 21.0, and 30.0 μg/mL sugiol and incubated for 24, 48, and 72 h. As a positive control, cells were treated with 2.0 μM H₂O₂ for the same time. After washing and resuspending the cells in PBS, DPPP (1,3-Bis(diphenylphosphino)propane - Sigma-Aldrich®) was added at a final concentration of 50 μM. The samples were incubated for 15 min at 25°C and the fluorescence measured on a Victor X3 fluorometer at λ_ex = 355 nm and λ_em = 460 nm. DPPP is a non-fluorescent molecule which reacts with fatty acids, phosphatidylcholine, and triacylglycerol hydroperoxides, resulting in a highly fluorescent molecule (Akasaka et al., 1987 (link); Takahashi et al., 2001 (link)).

2,5-dibromo-3-hexylthiophene ≥97%, TCI (Tokyo, Japan), [1,3-bis(diphenylphosphino)-propane]dichloronickel(II) (Ni(dppp)Cl₂, ≥97% Sigma Aldrich, St. Louis, MO, USA), vinylmagnesium bromide (0.7 M solution in tetrahydrofuran, Acros Organics (Geel, Belgium)), tert-butylmagnesium chloride (2 M solution in diethyl ether, Sigma Aldrich), anhydrous tetrahydrofuran (99.9%, Acros organics) distilled before use, poly(methylhydrosiloxane) trimethylsilyl terminated (average M_n = 390, Sigma Aldrich (Saint Louis, MO, USA)), poly(ethylene glycol) methyl ether methacrylate (average M_n = 500, M_n = 950 and M_n = 300, Sigma Aldrich), poly(ethylene glycol) methacrylate (average M_n = 500, Sigma Aldrich), platinum(0)-1,3-divinyl-1,1,3,3-tetramethyldisiloxane (PTDD, Karstedt’s catalyst) (complex solution in xylene, Pt ~ 2%, Sigma Aldrich), anhydrous toluene (99.85%, Acros Organics).