Tc100 medium

The IPL-LD 65Y cell line (Lymantria dispar) was obtained from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Braunschweig, Germany, No. ACC 181) and was maintained for routine culture in TC-100 medium (Lonza, Basel, Switzerland) with 11% fetal calf serum (FCS, PromoCell, Heidelberg, Germany). The cells were seeded with an initial concentration of 2E+05 cells per ml in tissue culture flasks (Greiner bio-one, Frickenhausen, Germany) and incubated at 27°C in a cooling incubator (Thermo Fisher Scientific, Schwerte, Germany). Cells were routinely passaged every seventh day.
Several commercially available substances were used in the screening assay. Fumagillin, surfactin, paromomycin, and metronidazole were obtained from Sigma-Aldrich (Taufkirchen, Germany); quinine, ornidazole, albendazole, tinidazole, clioquinol, and dimethylsulfoxid (DMSO) were obtained from VWR International (Darmstadt, Germany).

Larvae of Spodoptera exigua were obtained from a healthy laboratory colony maintained on a semi-synthetic diet [23 (link)] at 25 ± 1 °C. Sf9 cells (ThermoFisher Scientific, Waltham, MA, USA) were maintained in TC100 medium (Lonza Bioscience, Cologne, Germany) supplemented with 10% fetal calf serum (Lonza Bioscience, Cologne, Germany) at 28 ± 1 °C [24 ]. For the construction of the recombinant virus expressing the enhancin gene, the Bac-to-Bac^TM Baculovirus Expression System was used (ThermoFisher Scientific, Waltham, MA, USA). The enhancin gene was amplified from Trichoplusia ni granulovirus (TnGV) [16 (link)], whereas the polyhedrin gene was amplified from the AcMNPV C6 clone, the type species of the Alphabaculovirus genus [25 (link)]. All viruses were obtained from the virus collection of the Microbial Bioinsecticides group at the Universidad Pública de Navarra.