Journal: PLoS Pathogens

Article Title: Antiviral RNA interference in disease vector (Asian longhorned) ticks

doi: 10.1371/journal.ppat.1010119

Figure Lengend Snippet: (A) Phylogenetic analysis of Dicer-like proteins. Maximum-likelihood (ML) trees were constructed using MEGA-X software with 1000 bootstrap based on the protein sequences of each Dicer-like member. H . longicornis Dicer-like proteins were highlighted. C . elegans NP_498761.2 was set as outgroup to root the tree. (B) Conserved domain structures of Dicer-like proteins from D . melanogaster , I . scapularis and H . longicornis predicted by SMART database. Domains are indicated by colored boxes. The scale bars at the bottom represent the length of proteins in amino acid (aa). (C) Comparison of conserved amino acids in Dicer2-like proteins. (D and E) In vitro dicing of a synthetic 200 bp dsRNA by ectopically expressing FLAG-tagged EGFP (as a control), DCL2, and DCL1 immune-precipitated from S2 cells (D) or NoDice 293T cells (E). Western blotting detection of input and immune-precipitated Flag-tagged EGFP, DCL2, and DCL1. The Dicer substrate and product small RNAs were detected by 3% agarose gel with GelRed staining. Endogenous β-actin as a loading control.

Article Snippet: The FLAG-tagged proteins were retrieved using FLAG beads (GenScript) according to the manufacturer’s instructions.

Techniques: Construct, Software, Comparison, In Vitro, Expressing, Control, Western Blot, Agarose Gel Electrophoresis, Staining