Figure 3
From: Drosophila Dullard functions as a Mad phosphatase to terminate BMP signaling
Dullard physically interacts with Mad.
(a) S2R+ cells were co-transfected with Tkv-Flag and Dullard. Tkv-Flag was immunoprecipitated (IP) with anti-flag beads and then subjected to western blotting to evaluate if Tkv and Dullard interact. Results demonstrate Tkv and Dullard do not physically interact. (b) S2R+ cells were co-transfected with Flag-Mad and Dullard. Flag-Mad was immunoprecipitated (IP) with anti-flag beads and then subjected to western blotting to evaluate if Mad and Dullard interact. Results demonstrate Dullard and Mad proteins do physically interact. (c) Treatment of Flag-Mad lysates with calf intestinal phosphatase dephosphorylated Mad (shown by using pMadS212 antibody) compared to untreated lysates, compare lanes 3 and 4. Dephosphorylated Mad failed to bind Dullard compared to phosphorylated Mad, compare lanes 3 and 4. (d) S2R+ cells were co-transfected with Flag-Mad and Dullard-D66E. Flag-Mad was immunoprecipitated (IP) with anti-flag beads and then subjected to western blotting to investigate if both proteins interact. Results demonstrate Dullard-D66E and Flag-Mad proteins do physically interact. All western blots presented were repeated at least 2 times.
