Time-dependent transcript knockdown under constant exposure to long double-stranded (ds)RNA.

A, knockdown of virulence gene target transcripts after 72 h exposure to long dsRNA (CTRL, negative control; FABP, fatty acid binding protein; LAP, leucine aminopeptidase; μGST, μ-class glutathione transferase; ωGST, ω-class glutathione transferase; CatB, cathepsin B; CatL, cathepsin L; σGST, σ class glutathione transferase); B–D, time-dependent knockdown of cathepsin B (B), cathepsin L (C), σ-class glutathione transferase (D). Square brackets e.g. [CatB] represent qPCR amplicon. Target ΔΔCt (Y-axes) represents ratio of abundance of target transcript to a reference gene (glyceraldehyde phosphate dehydrogenase, GAPDH), in treated samples, relative to the abundance of those transcripts in untreated samples. Statistical significances are indicated relative to effects of negative control dsRNA (dsCTRL, complementary to neomycin phosphotransferase). dsCTRL treatments were performed in parallel with all experimental treatments. Experiments were repeated ≥4 times, employing 20–30 flukes per replicate. *, P<0.05; **, P<0.01; ***, P<0.001. Symbols represent mean±SEM.