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. 2001 Aug 6;154(3):631–644. doi: 10.1083/jcb.200106049

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Copyright © 2001, The Rockefeller University Press

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Figure 7. — EEA1 exclusion reduces LBPA accumulation on latex beads. (A) Quantitation of LBPA colocalization with latex bead phagosomes (30 min after infection) in cells injected with anti-hVPS34, anti-EEA1, control rabbit IgG, and uninjected control cells (n = 1,347 phagosomes). The data are means ± SE of three separate experiments. *P = 0.0001. (B) Quantitation of EEA1 colocalization with latex bead phagosomes (30 min after infection) in cells injected with anti-hVPS34, control rabbit IgG, and uninjected control cells (n = 601 phagosomes). The data are means ± SE of three separate experiments. *P < 0.001. (C) M. tuberculosis ManLAM but not its precursor PIM reduces EEA1 recruitment to the phagosome. Epifluorescence microscopy analysis of the effect of coating of latex beads with mycobacterial phosphoinositides on EEA1 recruitment. Latex beads were coated with purified M. tuberculosis H37Rv ManLAM or PIM and phagocytosed by J774 cells for 20 min. Quantitation of EEA1 colocalization with phagosomes (n = 1,321 phagosomes). Black bars, control or lipid-coated beads (as indicated) not treated with Triton X-100; hatched bars, control or lipid-coated beads (as indicated) extracted with Triton X-100 before infection. The data are means ± SE of three separate experiments. **P = 0.002. M. tuberculosis glycosylated phosphoinositides do not affect Syntaxin 8 acquisition by latex bead phagosomes. Epifluorescence microscopy analysis of Syntaxin 8 accumulation by phagosomes containing control uncoated latex beads or beads coated with mycobacterial phosphoinositides. Latex beads were coated with ManLAM or PIM and phagocytosed by J774 cells for 20 min. White bars, quantitation of Syntaxin 8 colocalization with phagosomes (n = 432 phagosomes). The data are means ± SE of three separate experiments. (D) M. tuberculosis ManLAM reduces LBPA recruitment to the phagosome. Epifluorescence microscopy analysis of the effect of coating of latex beads with ManLAM on LBPA accumulation. Quantitation of LBPA colocalization with phagosomes after 30 min of phagocytosis (n = 506 phagosomes). The data are means ± SE of three separate experiments. *P = 0.0001.