Figure 5

Analysis of association of PI3-kinase with transduced TAT-HA peptides Osteoclasts were transduced with TAT-peptides as indicated below the panels A and C. Panel A: Western analysis using an antibody to PI3-kinase. Lysates were immunoprecipitated with antibodies to HA and actin (lanes 1–6). Actin antibody was used as an internal control for immunoprecipitation. Immunoprecipitation with a non-immune serum (NI) is shown in lane 7. Panel B: Western analysis with an actin antibody. Panel C. Detection of association of PI3-kinase with endogenous gelsolin. Lysates were immunoprecipitated with a monoclonal antibody to gelsolin (lanes 2–7) or non-immune serum (NI; lane 1). Panel D: Western analysis using an antibody to gelsolin. To detect the levels of gelsolin, the immunoblot shown in panel C was stripped and blotted with an antibody to gelsolin. PI3-kinase, actin, gelsolin, and IgG heavy chain (IgG HC) protein bands are indicated by arrows. The results shown are representative of three independent osteoclast preparations and experiments.