Figure 6.

Mapping the interaction between ACT and p300. (A) Reciprocal endogenous Co-IP showing that ACT physiologically associates with p300 in vivo in the round spermatids. (B) ACT interacts with p300 through its first LIM domain. GFP tagged ACT full-length and deletion mutants harboring various number of LIM domains were constructed as indicated. Co-IP conducted using Flag-tagged p300, followed by western blot using GFP antibody against GFP-tagged ACT fragments. (C) p300 interacts with ACT through its GBD domain at the C-terminal. GST-pulldown was performed using GST-tagged p300 deletion mutants, followed by Western blot using the GFP antibody against the full-length ACT. Five deletion mutants of p300 constructs were constructed as described previously. Black arrowheads point to the expected protein size as predicted.