Schneider s drosophila medium

S2 cells were maintained at 25°C in Schneider's Drosophila Medium (Biowest) supplemented with 10% fetal bovine serum (Biochrom AG), 2 mM L-glutamine (PAA) and 1% penicillin/streptomycin (PAA). Transfections of S2 cells were done with Cellfectin II Reagent (Invitrogen) according to the manufacturer's protocol. For stable transfection pCo-Hygro was added to the transfection mixture and afterwards cells were selected with 200 µg/ml Hygromycin (InvivoGen). Protein expression was induced by addition of 0.7 mM CuSO₄ for about 20 h followed by cell lysis.

Leishmania infantum (MCAN/ES/92/BCN-83) was used in this experiment. Parasites were cultured in biphasic Novy-MacNeal-Nicolle (NNN) medium and sub-passaged in Schneider’s Drosophila medium (Biowest^®, Riverside, MO, USA) supplemented with 20% of fetal bovine serum ((FBS) Biowest^®, Riverside, MO, USA), 100 U/mL of penicillin (Life Technologies^TM, Carlsbad, CA, USA), and 1% of filtrated human urine from a healthy donor.

Trypanosoma brucei 29-13 was grown at 27 °C in SDM-79 (Thermo Fisher Scientific, Waltham, USA) supplemented with 10% (v/v) fetal bovine serum (Biosera, Cholet, France), 2-μg/mL hemin (Merck, Darmstadt, Germany), 50 U/mL penicillin, and 50-μg/mL streptomycin (both from Biowest, Nuaillé, France). Sergeia podlipaevi CER4 was grown at 23 °C in BHI medium (VWR/Avantor, Radnor, USA) supplemented as above. Paratrypanosoma sp. EC233, B. ayalai B08-376, V. ingenoplastis CP21, W. raviniae ECU-09, A. deanei CT-IOC-044, J. drosophilae Finn02, P. serpens 9T, H. tarakana OSR18, O. modryi Fi15, B. nonstop P57, N. esmeraldas E262, L. mexicana MNYC/BZ/62/M379, L. seymouri ATCC 30220, L. pyrrhocoris H10, C. brevicula S14, and C. thermophila CT-IOC-054 were grown at 23 °C in Schneider's Drosophila medium (Biowest) supplemented as above. Cells were collected in the exponential phase of growth (1 to 2 × 10⁷ cells/mL) for all experiments. Species identity was confirmed as in Yurchenko et al. (2016) (link).