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Fig. 8. KIN17 protein binds to RNA. (A) RNA homopolymer binding assay. Binding to agarose beads coupled to poly(A), poly(C), poly(G) and poly(U) is shown for wild-type mouse KIN17 protein, modified proteins KIN17₃₃₅KOW and KIN17₁₆₀ translated in a rabbit reticulocyte lysate. The binding of human KIN17 protein expressed in insect cells to RNA homopolymers and detected by western blot is also shown. A 1:3 ratio of the input protein is shown in each case. The binding of protein to homopolymers was evaluated by radioactivity detection using STORM phosphoimager and immunodetection with the exception of KIN17₁₆₀ protein that was not recognized by the mAb. Salt sensitivity of the binding was tested in buffers containing 0.1, 0.2, 0.5 and 1 M NaCl. (B) Northwestern binding assay using a [³²P]-labelled poly(G) or (C) [³²P]-labelled RNA probed against the proteins blotted on a nitrocellulose membrane. PSMW, prestained molecular weight markers, 1 µg of each protein was loaded.

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