American Society of Hematology

Fig. 3.

$Fig. 3. Distribution patterns of LacCer, transducer molecules, and NADPH oxidase components in fractions separated by sucrose density gradient centrifugation. / (A) Neutrophils, undifferentiated, and DMSO-treated HL-60 cells were lysed in the Triton X-100–contained lysis buffer, homogenized, and subjected to sucrose density gradient centrifugation. A series of 1 mL fractions were taken starting from the top and subjected to HPTLC (LacCer) or SDS-PAGE/immunoblotting (transducer molecules, NADPH oxidase components, and actin), respectively. DIM and high-density in undifferentiated and DMSO-treated HL-60 cells represent the mixture of fractions 5 and 6 and the mixture of fractions 9 to 12, respectively. (B) Neutrophils, undifferentiated and DMSO-treated HL-60 cells, and human umbilical vein endotheial cells (Endothelial cells) were lysed in the lysis buffer B, and the postnuclear supernatants of these cells were subjected to SDS-PAGE/immunoblotting with anticaveolin IgG.$

Distribution patterns of LacCer, transducer molecules, and NADPH oxidase components in fractions separated by sucrose density gradient centrifugation.

(A) Neutrophils, undifferentiated, and DMSO-treated HL-60 cells were lysed in the Triton X-100–contained lysis buffer, homogenized, and subjected to sucrose density gradient centrifugation. A series of 1 mL fractions were taken starting from the top and subjected to HPTLC (LacCer) or SDS-PAGE/immunoblotting (transducer molecules, NADPH oxidase components, and actin), respectively. DIM and high-density in undifferentiated and DMSO-treated HL-60 cells represent the mixture of fractions 5 and 6 and the mixture of fractions 9 to 12, respectively. (B) Neutrophils, undifferentiated and DMSO-treated HL-60 cells, and human umbilical vein endotheial cells (Endothelial cells) were lysed in the lysis buffer B, and the postnuclear supernatants of these cells were subjected to SDS-PAGE/immunoblotting with anticaveolin IgG.

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