Products/Services Used | Details | Operation |
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Proteins, Expression, Isolation and Analysis> | Anti-myc antibody (1:10,000 dilution, cat no. A00704, GenScript) while anti-AGO1, anti-AGO2 and anti-AGO4 (1:1000 dilution) were used for western blotting. AGO1, AGO2 and AGO4 antibodies were mcklh conjugated and raised against VRKRRTDAPSEGGEGC, NRGQGRGEQQC and CRELKKRNPNENGEFE peptides (Havecker et al., 2010) respectively, from Genscript (www.genscript.com)...Real-time PCR was performed using SYBR Green (Genscript) and primers are listed in Table S1. | Get A Quote |
The Cucumber mosaic virus (CMV) suppressor 2b co-localizes with AGO4 in cytoplasmic and nuclear fractions of Arabidopsis thaliana Biochemical fractionation of A. thaliana cellular extracts revealed that 2b and AGO4 coexist in multiple size exclusions. 2b transgenic A. thaliana exhibited an enhanced accumulation of 24nt siRNAs from flowering wageningen (FWA) and other heterochromatic loci. These plants also exhibited hypo-methylation of an endogenous- as well as transgene-FWA promoter at non-CG sites. In corroboration, both transgenic 2b and CMV infection affected the regulation of transposons which mimics the ago4 phenotype. In conclusion, 2b perturbs plant defense by interfering with AGO4-regulated transcr... More