Fig. 5: Nemacol-1 synergistically kills C. elegans in combination with Ivermectin.

a C. elegans viability over a double dose response matrix of Nemacol-1 and Ivermectin reporting the means of the control-normalized fraction of animals alive in treatment wells of 3-day growth assays (see 'Methods'). ~20 C. elegans L1s were added to wells containing the indicated condition with 0.6% DMSO and an HB101 E. coli food source in nematode growth medium. Data are from three biological replicates (N = 3) conducted in technical duplicate (n = 2). Significant differences of the EC50s relative to the ivermectin-only dose response are calculated using extra sum-of-squares F test. The respective p values are as follows: 2.8E-03 (3.8 µM Nemacol), 1.8E-03 (7.5 µM Nemacol), 1.6E-07 (15 µM Nemacol), 7.0E-08 (30 µM Nemacol), 1.4E-06 (60 µM Nemacol), and 7.5E-09 (e245 without Nemacol). The red and black asterisks indicates P < 0.01 and P = 0.001, respectively. b Zero Interaction Potency (ZIP) synergy score plot of the Ivermectin + Nemacol-1 double-dose response interaction generated using the SynergyFinder2.0 server⁴². c Dose-response curves from (a) highlighting the EC50 shifts of Ivermectin killing of wild type C. elegans in comparisons of no Nemacol treatment (black line, corresponding the values in the black box in (a)) versus 30 μM Nemacol-1 (orange line, corresponding the values in the orange box in (a)) (P = E-15), or no Nemacol treatment (black line) versus unc-17(e245) mutants (red line, corresponding the values in the red box in (a)) (P = 5.0E-15) reporting the SEM. P values are calculated using a two-sided extra sum-of-squares F test (see 'Methods'). d H. contortus control-normalized motility over a dose-response of Nemacol-53 showing the mean of three biological replicates (N = 3) and reporting the SEM (see 'Methods'). Four-parameter curves were fit in GraphPad 9.3.1 and used to calculate EC30 and EC50 values with symmetrical 95% confidence intervals. e H. contortus control-normalized motility over a dose-response of Ivermectin dose-response + /– an EC30 concentration of Nemacol-53 (26.4 µM) showing the mean of three biological replicates (N = 3) and reporting the SEM (see 'Methods'). Four-parameter curves were fit in GraphPad 9.3.1 and the associated P value (P = 0.0019) was calculated using a two-sided extra sum-of-squares F test comparing + /− 26.4 µM Nemacol-53 curves.