Projected cross-sectional images were reconstructed with maximum speed software. The intensity analyzed Of its fluorescence in the z-axis with ImageJ. Analysis of five repr Shown with representative Danoprevir ITMN-191 cells. Values of fluorescence Th projection cross-sectional side view images SHIP1 F Staining of the cells were analyzed HL-60 in suspension or by adhesion to ImageJ plotted against the lower surface Surface of the touch substrate medium of the cell, and an upper portion of the cell bound. n � �� �, * p 0 20 40 60 80 100 120 140 160 0 20 40 60 80 100 120 Mobile Cell2 a cell 3 cell 4 cell 5 0 20 40 60 80 100 120 0 20 40 60 80 100 mobile phone Cell2 a cell 3 cell 4 cell 5 0 2 4 6 8 10 12 A Susp Susp Adh Adh SHIP1 PD0325901 IP: SHIP1 4G10 No fMLP with fMLP B Susp Adh SHIP1 IP: SHIP1 b3 integrin FAK Lyn remote R & D intensity t in the z-axis at the bottom of the intensity t Relative greater than the distance from the axis E z * C bottom top SHIP1% PtdInsP3 actin dephosphorylation 0 20 40 60 80 100 120 in the lower center of the upper Unlk fertilization adherent SHIP1 intensity F t * 1224 | S. Mondal et al . Molecular biology of the cell, the cell migration process includes Zelladh Commission, indicated the involvement of integrin lead to the formation of binding sites on PtdInsP3. Above the production of Strength PtdInsP3 the bottom of a cell migration k nnte Required with the production of the front �� osterior PtdInsP3 gradient for cell migration. St Ren Our observations show that SHIP1 plays a role Role in the metabolism PtdInsP3 on the gel Walls of Zelladh Away mission.
M can Shortcomings in chemotaxis in SHIP1 � eutrophils by reducing the Zelladh recession SHIP1 be saved EUR eutrophils show strong adversely Chtigt migration toward a source of attractant. We then business protected That if chemotaxis due to excessive cell-adhesion recession Adversely Chtigt is, we may use the chemotaxis by reducing the surface Surface to improve adhesion. To resolve this problem, we examined neutrophil chemotaxis toward fMLP with EZ-cab scanning adhesion, we transfected Akt-PH-EGFP in neutrophils to PtdInsP3 see �� tdInsP2 wild-type and SHIP1 eutrophils �. They lie the cells on a surface impact area, coated with fibronectin. The cells were fixed and using high res Send confocal microscopy of cutting. SHIP1 � eutrophils had a significant accumulation of Akt-PH-EGFP in the cell cortex.
Of interest, disclose side view projection of images in cross section, that mission may need during the Adh, Akt-PH-EGFP on the plasma membrane of neutrophils wild type, but is in SHIP1 eutrophils �, there is a strong enrichment at the interface cell ubstratum. Since the loss SHIP1 prevent the formation of PtdInsP2 PtdInsP3 would enrichment was act-PH-EGFP formed primarily on an h Higher level of PtdInsP3 at the point of attachment. High concentrations of PtdInsP3 are indeed the decisive factor for the increased Hte Adh Sion of cells and activation of Akt in the Zelladh Sion in SHIP1 eutrophils �. These results suggest that even during the w pro-FIG 4: Adhesion-mediated signal transmission is being strengthened in PtdInsP3 SHIP1 EUR eutrophils verst.
Wild-type or SHIP1 � �o r wild-type PTEN or � eutrophils were either unstimulated or stimulated with 1 M fMLP μ suspended or just to keep the coating on a surface Surface with fibronectin for 15 min cells , and not adhering kidnapped min and then stimulated with fMLP for 2. The phospho-Akt were analyzed in cell lysates. Total Akt was used as a contr The load. Wild-type or SHIP1 � eutrophils were transfected with Akt-PH-EGFP and you lie keep it on a surface surface coated with fibronectin. The cells were fixed and with the help of confocal microscopy, images were reconstructed cross-sectional side view projection heavy accumulation of Akt-PH-EGFP in the cell interface ubstratum shows SHIP1 eutrophils �. The intensity analyzed Of its fluorescence in the z-axis with ImageJ. Analysis of four repr Shown with representative cells for each genotype. Values of fluorescence intensity Th edge of the side view cross-sectional images of the wild-type and SHIP1 � Ellen expressed Akt-PH-EGFP were analyzed by ImageJ Plott