Figure 2
From: Polyphosphoinositides-dependent regulation of the osteoclast actin cytoskeleton and bone resorption
Analysis of PtdIns P 2 association with the transduced protein. Panel A. Western analysis with an anti-PtdIns P2 antibody: Treatments are as indicated below the figure. Lysates were immunoprecipitated with an antibody to HA (lanes 2–9) or non-immune serum (NI; lane 1). The immunoprecipitates were subjected to 8% SDS-PAGE (lanes 1 to 3) or 15% SDS-PAGE (lanes 4 to 9) and Western analyzed with an antibody to PtdIns P2. PtdIns P2 binding to the transduced peptides is indicated with arrowheads. Arrows indicate IgG heavy chain (IgG HC). The results shown are representative of three independent experiments. Panel B. TLC analysis of the phospholipids associated with the anti-HA immunoprecipitates: 32P-labeled osteoclasts were subjected to various treatments as indicated in the figure. Osteoclast lysates made from these cells were subjected to immunoprecipitation with an antibody to HA. 32P-labeled phosphoinositides associated with anti-HA immunoprecipitates were analyzed by TLC. Arrows on the left side of the figure indicate the migration of PtdIns P2 and PtdIns P3. The results shown are representative of three independent osteoclast preparations and experiments.