American Society of Hematology

Figure 1.

$Figure 1. Detection of PSGL-1 and L-selectin in lipid rafts. (A) Detergent-insoluble fractions of KG1 or Jurkat cell lysates were isolated at the 5%/30% interface of a discontinous sucrose gradient. Lipid rafts containing fractions isolated from KG1 cells (A, top panel) or Jurkat cells (B, top panel) were revealed by dot-blot analysis, using biotinylated B-subunit of cholera toxin (A and B, top panels). After electrophoresis on a 7.5% SDS polyacrylamide gel, gradient fractions isolated from KG1 cells (A, bottom panel), Jurkat cells (B, bottom panel), or neutrophils (C and D) were examined for the presence of PSGL-1 (A and C) or L-selectin (B and D) by immunoblotting. (E) Histogram of PSGL-1 levels in fractions 1 to 10 (mean ± SEM, n = 3). (F) Histogram of L-selectin levels in fractions 1 to 10 (mean ± SEM, n = 4).$

Detection of PSGL-1 and L-selectin in lipid rafts. (A) Detergent-insoluble fractions of KG1 or Jurkat cell lysates were isolated at the 5%/30% interface of a discontinous sucrose gradient. Lipid rafts containing fractions isolated from KG1 cells (A, top panel) or Jurkat cells (B, top panel) were revealed by dot-blot analysis, using biotinylated B-subunit of cholera toxin (A and B, top panels). After electrophoresis on a 7.5% SDS polyacrylamide gel, gradient fractions isolated from KG1 cells (A, bottom panel), Jurkat cells (B, bottom panel), or neutrophils (C and D) were examined for the presence of PSGL-1 (A and C) or L-selectin (B and D) by immunoblotting. (E) Histogram of PSGL-1 levels in fractions 1 to 10 (mean ± SEM, n = 3). (F) Histogram of L-selectin levels in fractions 1 to 10 (mean ± SEM, n = 4).

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