American Society of Hematology

Figure 2

$Figure 2. B-NHL rafts contain a predominant tyrosine phosphorylated component of 80 kDa. (A) Subcellular fractionation of TX-100 lysates as in Figure 1; 10 μL of each gradient fraction (as in Figure 1A) was probed with the antiphosphotyrosine 4G10 mAb. (B) Tyrosine phosphorylated PAG is part of the 80-kDa raft component. Raft and nonraft gradient fractions from the indicated cell lines containing each equivalent amounts of anti-PAG-reactive protein. R indicates fraction 3-5 (raft) proteins; NR, fraction 10-11 (nonraft) proteins. First row, WB with antiphosphotyrosine 4G10 antibody; second row, WB with antibody against peptide KEISAMpYSS, recognizing pY317 in PAG; third row, WB with anti-PAG antibody MEM-255. (C) Doubly tyrosine phosphorylated Lyn is part of the 80-kDa component in selected B-NHL rafts. Raft (R) and nonraft (NR) gradient fractions from the indicated cell lines containing each equivalent amounts of Lyn. First row, WB with anti-pY418 Src antibody; second row, WB with anti-pY505 Lck antibody; third row, WB with anti-Lyn antibody. Open arrowheads indicate the 80-kDa, antiphospho-Lyn-reactive component. Gradients and IPs were carried out 3 times for all cell lines, and representative experiments are shown.$

B-NHL rafts contain a predominant tyrosine phosphorylated component of 80 kDa. (A) Subcellular fractionation of TX-100 lysates as in Figure 1; 10 μL of each gradient fraction (as in Figure 1A) was probed with the antiphosphotyrosine 4G10 mAb. (B) Tyrosine phosphorylated PAG is part of the 80-kDa raft component. Raft and nonraft gradient fractions from the indicated cell lines containing each equivalent amounts of anti-PAG-reactive protein. R indicates fraction 3-5 (raft) proteins; NR, fraction 10-11 (nonraft) proteins. First row, WB with antiphosphotyrosine 4G10 antibody; second row, WB with antibody against peptide KEISAMpYSS, recognizing pY317 in PAG; third row, WB with anti-PAG antibody MEM-255. (C) Doubly tyrosine phosphorylated Lyn is part of the 80-kDa component in selected B-NHL rafts. Raft (R) and nonraft (NR) gradient fractions from the indicated cell lines containing each equivalent amounts of Lyn. First row, WB with anti-pY418 Src antibody; second row, WB with anti-pY505 Lck antibody; third row, WB with anti-Lyn antibody. Open arrowheads indicate the 80-kDa, antiphospho-Lyn-reactive component. Gradients and IPs were carried out 3 times for all cell lines, and representative experiments are shown.

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