American Society of Hematology

Figure 1

$Figure 1. PAG and Lyn codistribute in detergent-resistant raft membranes of B-NHLs. (A) Subcellular fractionation of TX-100 lysates. TX-100 lysates (50 × 106 cells) of the indicated cell line loaded on sucrose gradient, ultracentrifuged to equilibrium, and 10 μL of each fraction was probed by WB with anti-PAG (MEM-255), anti-Lyn and anti-Csk. Fractions 1 to 11, from top to bottom. p: pellet of the equilibrium gradient containing nuclei and cytoskeleton. Dotted lines indicate the raft-containing fractions 3-5. (B) Sedimentability of the TX-100-resistant material. An aliquot (10 μL) of each gradient fraction was diluted 100-fold and ultracentrifuged (100 000g for 1 hour). The sedimented material probed by WB and raft fractions between dotted lines, as in panel A. Gradient fractionation was carried out at least twice for all cell lines and more than 5 times for Raji, DoHH2, SUDHL6, ALK+, and L428 lines.$

PAG and Lyn codistribute in detergent-resistant raft membranes of B-NHLs. (A) Subcellular fractionation of TX-100 lysates. TX-100 lysates (50 × 10⁶ cells) of the indicated cell line loaded on sucrose gradient, ultracentrifuged to equilibrium, and 10 μL of each fraction was probed by WB with anti-PAG (MEM-255), anti-Lyn and anti-Csk. Fractions 1 to 11, from top to bottom. p: pellet of the equilibrium gradient containing nuclei and cytoskeleton. Dotted lines indicate the raft-containing fractions 3-5. (B) Sedimentability of the TX-100-resistant material. An aliquot (10 μL) of each gradient fraction was diluted 100-fold and ultracentrifuged (100 000g for 1 hour). The sedimented material probed by WB and raft fractions between dotted lines, as in panel A. Gradient fractionation was carried out at least twice for all cell lines and more than 5 times for Raji, DoHH2, SUDHL6, ALK⁺, and L428 lines.

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