Figure 2
A) Northern blot analysis of AMSH-2 in different human tissues and cell lines. Each lane contains 2 μg of poly A+ mRNA. The top panels show the result of hybridization of the membranes with a specific probe for AMSH-2 mRNA. The lower panels show β-actin reprobing of the membranes. The sizes of the transcripts are indicated in kb on the left. B) Expression of AMSH-2 protein. Cell lysates from metabolically labeled 293T and HepG2 cells were immunoprecipitated with preimmune (PI) and AMSH-2 antiserum (I). A specific band for AMSH-2 can be observed in the lanes immunoprecipitated with AMSH-2 antiserum (indicated by an arrow). C) Expression of AMSH-2 Myc construct. 293T cells were transfected with the indicated constructs and metabolically labeled with S35. Cell lysates were immunoprecipitated with 9E10 monoclonal antibody and after SDS-polyacrylamide gel separation and blotting, the membrane was exposed. The arrow points a specific band corresponding to the Myc tagged AMSH-2 protein.