To assess expression of EGFP, eggs were scored as positive when >20% of the miracidium surface area was EGFP positive; representative EGFP-positive eggs are shown in panels C2 and C3. On day 15 following transfection, eggs were scored and counted for EGFP positivity in each of four biological replicates (~100 eggs/replicate) of KI and lsDNA-only eggs. About 25% of the eggs in the lsDNA-only group were scored as EGFP positive (19%–32%), whereas 75% of eggs in the KI group expressed EGFP (68%–79%); p < 0.001; two-tailed t = 69.87, df = 142; difference between means (EGFP-KI-only donor) ± SEM, 49.7 ± 0.7, 95% CI, 48.3–51.1.
