Figure 3.

Src251 is detergent-insoluble in osteoclasts. (A) Osteoclasts were differentiated from bone marrow in culture, then lysed gently in 0.5% Triton X-100 for varying times (2-min lysis is shown here; similar results were obtained at other time points) and the remaining detergent-insoluble fraction then lysed with RIPA. Src was immunoprecipitated from detergent-soluble and -insoluble fractions and analyzed by immunoblotting (Kaplan et al. 1994). (B) Detergent solubility of Pyk2 and AKT are not altered by expression of Src251. Osteoclasts were lysed as above, and lysates were immunoblotted for Pyk2 and AKT. Similar results were seen at 1-, 2-, and 4-min detergent solubility lysis conditions (1- and 4-min conditions are shown here). (S) Detergent-soluble fraction; (In) detergent-insoluble fraction.