Skip to main content
. 2011 Apr 28;12(6):581–586. doi: 10.1038/embor.2011.66

Figure 3.

Figure 3

RACK1 interacts with human Ago2 and miRNAs. (A) Human RACK1 binds to both Ago2 and miRNAs. RACK1 was immunoprecipitated with monoclonal RACK1 antibody (RACK1) and non-conjugated Protein A beads as a negative control. Input represents the equivalent of 4% of the total extract used for immunoprecipitation. The immunoprecipitate was tested for the presence of RACK1, hAgo2 and hAgo1 by western blotting and let-7 miRNA by northern hybridization. (B) RACK1 interaction with hAgo2 and the miRNA miR-21 is RNA independent. Immunoprecipitations were performed with mouse monoclonal antibodies raised against RACK1 and GFP (used as a negative control) as indicated. Input represents 10% of total lysate used for the immunoprecipitation. The immunoprecipitates were treated with RNase A for 1 h at 4°C. The supernatant (S) and the immunoprecipitate (B) samples, with or without RNase treatment, were tested for the presence of hAgo2, RACK1 and miR-21. GFP, green fluorescent protein; hAgo1/2, human Ago1/2; IP, immunoprecipitation; miRNA, microRNA; RACK, receptor for activated C-kinase; RNAi, RNA interference.