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Fig. 5. Osteoblastic cells contain detergent insoluble anx2. (A) Solublization of U2OS cells in 1% Triton X-100 results in a soluble fraction of anx2 (S), and an insoluble fraction of anx2 (P). (B) Cellular lysates were made from both SaOSLM2-anx2GFP-(1) and GFP-(2) expressing cells. Conditioned medium (CM) was also collected from these cells. (C) The membranous fraction following subcellular fractionation of SaOSLM2 cells was treated with 0.01% digitonin or the diluent methanol for 10 minutes. Membranes were resuspended and these insoluble membrane pellets (P) as well as the soluble wash (W) components were analyzed by immunoblot for the presence of anx2.





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