Figure 5. A phospho-mimetic form of DAZAP1 does not interact with DAZ.

(A) HEK-293 cells were transfected without (−) or with 2.5 μg of FLAG-DAZ plasmid DNA (+), 2.5 μg of DNA expressing unmodified HA–DAZAP1(155–407) (WT), HA–DAZAP1(155–407) in which Thr²⁶⁹ and Thr³¹⁵ had both been mutated to Ala (AA), HA–DAZAP1(155–407) in which Thr²⁶⁹ and Thr³¹⁵ had both been mutated to Asp (DD) or singly mutated forms of DAZAP1 in which either Thr²⁶⁹ or Thr³¹⁵ had been mutated to Asp (T269D and T315D). After 36 h, DAZAP1 was immunoprecipitated (IP) from 1 mg of cell extract protein with an anti-HA antibody and immunoblotted (IB) with anti-HA and anti-FLAG antibodies. (B) As described for (A), except that the cells were transfected with full-length forms of DAZAP1.