FIGURE 2.
Specificity of the interaction between FKBP38 and ANKMY2. A, extracts of HEK293T cells transiently transfected with expression vectors for 3×FLAG-tagged human FKBP38 or human FKBP52 (negative control) and for 2×HA-tagged human ANKMY2 were subjected to immunoprecipitation with anti-FLAG. The resulting precipitates, as well as a portion (2% of the input for immunoprecipitation (IP)) of the cell extracts, were subjected to immunoblot analysis (IB) with anti-HA, anti-FLAG, and anti-calnexin. B, extracts of HEK293T cells transiently transfected with expression vectors for 3×FLAG-tagged ZMYND proteins were subjected to immunoprecipitation with anti-FLAG. The resulting precipitates, as well as a portion (3% of the input for immunoprecipitation) of the cell extracts, were subjected to immunoblot analysis with anti-FKBP38, anti-FLAG, and anti-calnexin. C, His6-T7-tagged mouse FKBP38 was incubated with GST-tagged mouse ANKMY2 or GST (negative control), and the binding mixtures were then subjected to precipitation with glutathione-conjugated beads. The bead-bound proteins (pull-down), as well as a portion (10% of the input for precipitation) of the binding mixtures, were subjected to immunoblot analysis with anti-T7 and anti-GST.