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Fig. 5. HDAC1 binds p73{alpha} in vivo. (A) COS-1 cells were co-transfected with Flag-tagged p73{alpha}, or Flag-vector together with HA-tagged HDAC1 or HA vector, respectively. Cell lysates were reciprocally immunoprecipitated (IP) and immunoblotted (IB) with Flag antibody or HA antibody, as described in legend to Fig. 4. Cell lysates were immunoblotted with HA antibody (upper panel) or Flag-M2 antibody (lower panel). (B) GST-HDAC1 and HA-p73{alpha} fusion proteins were incubated in vitro and immunoprecipitated with GST antibody. Binding was detected using antibodies against HA and GST. (C) Flag-tagged p73{alpha} or its C-terminal deletion mutants were co-transfected with HA-tagged HDAC1 or vector, and the interaction was examined as described above. (D) Interaction between {Delta}Np73{alpha} and HA-tagged HDAC1 was examined as described in (A). In addition, anti-{Delta}Np73 was used for immunoprecipitation and immunoblotting.