Figure 5

In vivo RNAi in tick salivary glands. Unfed, adult female I. scapularis ticks were first micro-injected with either A) dsRNA-β-Actin or buffer or B) dsRNA Na⁺-K⁺-ATPase or buffer and then allowed to blood feed for 3–4 days on rabbits. Salivary glands were dissected from partially fed mock-injected and knockout ticks and pooled into two groups. Total RNA was extracted and cDNA produced. RT-PCR was conducted followed by gel electrophoresis. A) β-Actin transcript levels demonstrating the efficiency of RNAi in tick salivary glands, (Lane 1, Low DNA Mass™ Ladder, 2–3: Mock and gene silenced samples amplified with β-actin gene specific primers, 4–5: mock and gene silenced samples amplified with Cyclophilin A gene specific primers), B) Na⁺-K⁺-ATPase transcript levels (Lane 1, Low DNA Mass™ Ladder, 2–3: Mock and gene silenced samples amplified with Na⁺-K⁺-ATPase gene specific primers, 4–5: mock and gene silenced samples amplified with cyclophilin G gene specific primers). Low DNA Mass™ ladder (100–2000 bp; 2000 bp, 1200 bp, 800 bp, 400 bp, 200 bp, 100 bp) was used from Invitrogen.