Eukaryotic translation initiation factor 2C (eIF2C) proteins (argonaute family) influence RNA interference (RNAi) as components of the RNA-inducible silencing complex (RISC) or microRNA (miRNA)-containing ribonucleoprotein particle (miRNP). Small RNAs, including small interfering RNAs (siRNAs) and miRNAs, can silence target genes through mechanisms that utilize RISC or miRNP particles. eIF2C1 (argonaute 1, AGO1, eIF2C, GERP95, Q99) and Dicer1 play a coordinated role in siRNA-mediated gene silencing. eIF2C2 (Slicer, argonaute 2, AGO2, Q10) is a RISC component that can concentrate in cytoplasmic processing bodies (P-bodies) and catalyze mRNA cleavage. Mammalian P-bodies contain mRNAs and have an association with miRNA-induced translational silencing and siRNA-induced mRNA degradation. Additional eIF2C proteins include eIF2C3 (argonaute 3, AGO3), eIF2C4 (argonaute 4, AGO4) and meIF2c5 (mouse argonaute 5).
Background References
1. Hein M.Y., et al. 2015. A human interactome in three quantitative dimensions organized by stoichiometries and abundances. Cell 163:712-723.
2. Schurmann N., et al. 2013. Molecular dissection of human Argonaute proteins by DNA shuffling. Nat. Struct. Mol. Biol. 20:818-826.
Western blot analysis of AGO3 on different lysates with Rabbit anti-AGO3 antibody (ET1701-26) at 1/1,000 dilution.
Lane 1: HeLa cell lysate Lane 2: C6 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 97 kDa Observed band size: 97 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-26) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ICC staining AGO3 in N2A cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
ICC staining AGO3 in SHG-44 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
ICC staining AGO3 in F9 cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
ICC staining AGO3 in NCCIT cells (green). The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Immunohistochemical analysis of paraffin-embedded rat spinal cord tissue using anti-AGO3 antibody. Counter stained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-AGO3 antibody. Counter stained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-AGO3 antibody. Counter stained with hematoxylin.
Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-AGO3 antibody. Counter stained with hematoxylin.
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