FIG. 4.

Rep68 binds the C terminus of ANP32B but not ANP32A in vitro. (A) Schematic representation of functional domains of ANP32 proteins and of the mutant proteins used in panels B and C. LRR, leucine-rich repeats. (B) In vitro pull-down experiments. In vitro-transcribed-translated (IVT), [³⁵S]methionine-labeled ANP32 proteins were incubated with recombinant His-Rep68 bound on Ni-NTA beads. The levels of bound proteins were measured as the quantity of radioactivity that was retained on the Ni-NTA beads (as counted by PhosphorImager) and expressed as the percentage of the input. (C) Same as panel B, but deletion mutant forms of ANP32B were used in in vitro pull-down assays with His-Rep68 as indicated. The same experiments were independently performed at least three times, and superimposable results were obtained.