Systematic Identification of mRNAs Recruited to Argonaute 2 by Specific microRNAs and Corresponding Changes in Transcript Abundance
Figure 1
Ago2 association with Dicer and miRNAs.
(A) Strategy for the systematic isolation and identification of RNA associated with Ago2. (B) Immunopurification of FLAG protein purified from mock transfected cells (left) and FLAG-Ago2 transfected cells (right). A Dicer antibody (top) and FLAG antibody (bottom) were immuno-reactive with bands corresponding to the predicted molecular weight of Dicer (∼250 kD) and Ago2 (∼90 kD). (C) SYRO ruby protein stain of FLAG immunopurifications from mock transfected cells (left) and FLAG-Ago2 transfected cells (right). (D) SYBR gold nucleic acid stain of small RNAs (20–40 bp) isolated from whole cell lysate (left), FLAG immunopurification from FLAG-Ago2 transfected cells (middle) and FLAG purification from mock transfected cells (right). Brackets outline expected migration of nucleic acids ∼21 base pairs in length.