Journal: Viruses

Article Title: Regulation of RNA Interference Pathways in the Insect Vector Laodelphax striatellus by Viral Proteins of Rice Stripe Virus

doi: 10.3390/v13081591

Figure Lengend Snippet: Response of miRNA and siRNA pathways to RSV infection in planthoppers. ( A ) The relative RNA levels of RSV NP in the planthoppers after injection of RSV crude preparations from viruliferous planthoppers measured with quantitative real-time PCR (qPCR). Significant differences are indicated by different lowercase letters. ( B – I ) The relative transcript levels of Drosha , Pasha , Dicer1 , Ago1 , Dicer2 , Ago2 , Translin, and Trax in the miRNA and siRNA pathways during the infection process of RSV in the planthoppers were measured with qPCR. The crude preparations from nonviruliferous planthoppers were injected into the control groups (CK). The RNA or transcript levels of these genes were normalized to that of EF2 . The values represent the means ± SEs. NS, no significant difference. **, p < 0.01.

Article Snippet: The purified Translin-His served as the antigen to produce an anti-Translin rabbit polyclonal antibody at the Beijing Protein Institute Co., Ltd. (Beijing, China).

Techniques: Infection, Injection, Real-time Polymerase Chain Reaction, Control

Journal: Viruses

Article Title: Regulation of RNA Interference Pathways in the Insect Vector Laodelphax striatellus by Viral Proteins of Rice Stripe Virus

doi: 10.3390/v13081591

Figure Lengend Snippet: Western blot assay showing the protein levels of NP, Ago1, Ago2, and Translin in the planthoppers at 6 d after injection of RSV crude preparations. The crude preparations from nonviruliferous planthoppers were injected into the control group (CK). Three biological replicates were shown for each group. The monoclonal antibodies for NP, Ago1 and Ago2, and polyclonal antibody for Translin were used. The protein level of planthopper tubulin was revealed using the monoclonal anti-β-tubulin antibody as an internal reference. The relative grayscales of Ago1, Ago2, and Translin to that of tubulin were quantified and compared between the RSV-infected and CK groups. The values represent the means ± SEs. NS, no significant difference.

Article Snippet: The purified Translin-His served as the antigen to produce an anti-Translin rabbit polyclonal antibody at the Beijing Protein Institute Co., Ltd. (Beijing, China).

Techniques: Western Blot, Injection, Control, Bioprocessing, Infection

Journal: Viruses

Article Title: Regulation of RNA Interference Pathways in the Insect Vector Laodelphax striatellus by Viral Proteins of Rice Stripe Virus

doi: 10.3390/v13081591

Figure Lengend Snippet: Roles of miRNA and siRNA pathways in the control of RSV replication. ( A – C ) Effect of planthopper Ago1 , Ago2 , and Translin transcript levels, and viral NP and RNA3 levels at 6 d after injection of double-stranded RNAs (dsRNAs) with RSV crude preparations. The control group was injected with ds GFP -RNA and RSV crude preparations. ( D ) Effect of planthopper Translin transcript level and viral NP and RNA3 levels in the stable viruliferous strain at 6 d after injection of ds Translin -RNA. The control group was injected with ds GFP -RNA. The relative RNA levels of NP and RNA3 to that of EF2 and relative transcript levels of the three planthopper genes to that of EF2 were quantified with quantitative real-time PCR. The protein levels of NP and tubulin were measured using monoclonal anti-NP and anti-β-tubulin antibodies by western blot. Four biological replicates were shown for each group. The relative grayscale of NP to that of tubulin was quantified and compared between the gene interference groups and ds GFP -RNA injection groups. The values represent the means ± SEs. *, p < 0.05. **, p < 0.01. ***, p < 0.001. NS, no significant difference.

Article Snippet: The purified Translin-His served as the antigen to produce an anti-Translin rabbit polyclonal antibody at the Beijing Protein Institute Co., Ltd. (Beijing, China).

Techniques: Control, Injection, Real-time Polymerase Chain Reaction, Western Blot

Journal: Viruses

Article Title: Regulation of RNA Interference Pathways in the Insect Vector Laodelphax striatellus by Viral Proteins of Rice Stripe Virus

doi: 10.3390/v13081591

Figure Lengend Snippet: Regulation of RNAi pathways through insect protein-viral protein interactions. ( A ) Coimmunoprecipitation (Co-IP) assay demonstrating that the recombinant expressed NS2-His pulled down Ago2 from the crude extracts of viruliferous planthoppers. ( B ) Co-IP assay demonstrating that the recombinant expressed RdRp2-His pulled down Translin from the crude extracts of viruliferous planthoppers. Monoclonal antibodies against Ago2 and His and polyclonal antibodies against Translin were applied. IgG was used as a negative control. ( C ) Effect of planthopper Ago1 and Ago2 transcript levels and viral NS2 , NP, and RNA3 levels in the planthoppers at 6 d after injection of double-stranded RNAs (dsRNAs) of NS2 with RSV crude preparations. The control group was injected with ds GFP -RNA and RSV crude preparations. The relative transcript or RNA levels to that of EF2 were quantified with quantitative real-time PCR. The protein levels of NP and tubulin were measured using monoclonal anti-NP and anti-β-tubulin antibodies by western blot. Four biological replicates were shown for each group. The relative grayscale of NP to that of tubulin was quantified and compared between the gene interference group and ds GFP -RNA injection groups. The values represent the means ± SEs. *, p < 0.05. NS, no significant difference.

Article Snippet: The purified Translin-His served as the antigen to produce an anti-Translin rabbit polyclonal antibody at the Beijing Protein Institute Co., Ltd. (Beijing, China).

Techniques: Co-Immunoprecipitation Assay, Recombinant, Bioprocessing, Negative Control, Injection, Control, Real-time Polymerase Chain Reaction, Western Blot