Fig. 1
dsRNA-mediated knockdown of transcripts for I. scapularis genes of interest in ISE6 cells. Following transfection of ISE6 cells with 10 ng dsRNA for 60 h, total RNA was prepared from ~ 1 × 105 cells and cDNA was amplified via a two-step RT-PCR reaction. mRNA levels were normalized to I. scapularis β-actin and expressed relative to the percentage of pGEM control cDNA. Results show relative expression of ten I. scapularis genes of interest following knockdown (white bars) relative to the pGEM dsRNA negative control (gray bars). Error bars represent standard error of the mean (SEM). Statistical analysis was performed using an unpaired t-test between the negative pGEM control and dsRNA treatments for each gene of interest. Results represent 2–3 technical replicates (each with 2–3 machine replicates) and 2 biological replicates. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001. Abbreviations: FAH, fumarylacetoacetase (ISCW020196); ERP29, endoplasmic reticulum protein 29 (ISCW018425); ALDH, aldehyde dehydrogenase (ISCW015982); VNN, carbon-nitrogen hydrolase/vanin-like (ISCW004822); MDH2, malate dehydrogenase (ISCW003528); PARP, poly [ADP-ribose] polymerase (ISCW019519); CMPK, cytidine/uridine monophosphate kinase (ISCW012446); ACAT1, acetyl-CoA acetyltransferase (ISCW016117); Hypo195, hypothetical protein (ISCW011195); Hypo576, hypothetical protein (ISCW020576); pGEM, pGEM plasmid (negative control)