Figure 4

In vivo RNAi and expression of Dicer 1 like gene in tick synganglia. Unfed, adult female I. scapularis ticks were first micro-injected with either dsRNA-β-Actin or buffer or dsRNA Na⁺-K⁺-ATPase or irrelevant LacZ gene and then allowed to blood feed for 3–4 days on rabbits. Synganglia were dissected from partially fed mock-injected and gene silenced 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 synganglia, (Lane 1, AmpliSize Molecular Ruler, 2: buffer injected, 3: LacZ injected, 4: Actin-dsRNA injected, 5: Na⁺-K⁺-ATPase-dsRNA injected synganglia amplified with β-actin gene specific primers, B) Na⁺-K⁺-ATPase transcript levels demonstrating the effectiveness of RNAi in tick synganglia, (Lane 1, AmpliSize Molecular Ruler, 2: buffer injected, 3: LacZ injected, 4: Actin-dsRNA injected, 5: Na K ATPase-dsRNA injected synganglia amplified with Na+-K+-ATPase gene specific primers.