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. 2012 May 21;287(27):22450–22462. doi: 10.1074/jbc.M112.339663

FIGURE 1.

FIGURE 1.

Effect of M6P-IGF2R silencing on the lipid raft partitioning of uPAR. Control (shCTR)- and M6P-IGF2R-silenced (shM6P-IGF2R) TCL-598 cells were lysed by using the detergent Brij-58, and the cell lysates were subjected to sucrose gradient density centrifugation. Ten individual gradient fractions were taken gradually from the top (light fractions corresponding to detergent-resistant lipid rafts) to the bottom (heavy fractions corresponding to soluble proteins). The samples were analyzed by SDS-PAGE followed by immunoblotting (IB) with mAb specific to M6P-IGF2R (mAb MEM-238), β3 integrin (mAb VIPL-2), uPAR (mAb H2), pro-uPA (mAb U5), and CD59 (mAb MEM-43/5) as a control of lipid raft fractionation. Chemiluminescence was used for visualization. D1D2D3 points to the band corresponding to the full-length uPAR, and D2D3 points to the band corresponding to the truncated form. Results are representative for three independent experiments.