C. elegans Killing Assay with Curcumin

The C. elegans killing assay used was a modification of a previously described protocol (Beceiro et al., 2014 (link)). Briefly, non-infected nematodes (∼20–30) [fer-15(b26);fem-1(hc17)] were pipetted into 96-well plate containing M9 buffer and overnight curcumin (50 μg/ml) treated and untreated with A. baumannii and/or C. albicans cells. As a second dose curcumin was added to respective wells to make final concentration 50 μg/ml (total volume 300 μl). Nematodes were incubated at 25°C and viabilities were determined as previously described (Rajsekharan et al., 2018 (link)), by exposing them to LED or UV LED lights for 10–30 s using an iRiS^TM Digital Cell Imaging System (Logos BioSystems, South Korea). Three independent experiments (n = ∼20–30) were conducted.