0.4 cm electroporation cuvette

Knockdown of RNAs encoding SMDR2 (NCBI Acc. #L26287), SmMRP1 (NCBI Acc. #GU967672), ABCA4 (Smp_056290), ABCB6 (Smp_134890), and MRP7/ABCC10 (Smp_147250) was as described [47] (link), [81] (link). Briefly, following an overnight incubation in schistosome medium, adult worms (5 males plus 5 females) were placed in a 0.4 cm electroporation cuvette (USA Scientific, Ocala, FL) containing 50 µl siPORT (Life Technologies, Grand Island, NY) plus 3 µg of each of the siRNAs (IDT, Coralville, IA), either singly or in combination, targeting SMDR2, SmMRP1, MRP7, ABCA4, and ABCB6, or up to 15 µg luciferase siRNA (Life Technologies, Grand Island, NY; 3 µg per experimental siRNA used). The luciferase siRNA used for our control shows no significant similarity to any sequences from the S. mansoni gene database. siRNAs against the S. mansoni transporters were designed using the IDT SciTools RNAi Design server; sequences and targets are listed in Table 1. Worms were electroporated in this solution with a 20 ms square-wave pulse of 125 volts (BioRad Pulser XCell). Following electroporation, worms were incubated in schistosome medium for 2 days. They were then sorted into 2–3 males/female pairs per well in a 12-well plate, in which they were subsequently incubated in schistosome medium with carrier alone, or in medium plus PZQ (800 nM), as described above, and subsequently analyzed for motility.