Rapid Membrane Disruption by a Perforin-Like Protein Facilitates Parasite Exit from Host Cells

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Science  18 Dec 2008:

DOI: 10.1126/science.1165740

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  • Rapid Membrane Disruption by a Perforin-Like Protein Facilitates Parasite Exit from Host Cells
    Björn F. C. Kafsack, Janethe D. O. Pena, Isabelle Coppens, Sandeep Ravindran, John C. Boothroyd, Vern B. Carruthers

    Supporting Online Material

    This supplement contains:
    Materials and Methods
    Figs. S1 to S6
    References

    This file is in Adobe Acrobat PDF format.

    Other Supporting Online Material for this manuscript includes the following: (available at www.sciencemag.org/cgi/content/full/1165740/DC1)
    Movies S1 to S7

    Movie s1
    Phase contrast videomicroscopy of ionophore-induced egress by WT tachyzoites from infected fibroblasts at 30h post-infection. Time (h:min:sec) elapsed following calcium ionophore addition is shown. Note the short time between motility activation and egress from the PV. Also note the near synchrony of egress events.

    Movie s2
    Phase contrast videomicroscopy of ionophore-induced egress by plp1ko tachyzoites from an infected fibroblast at 30h post-infection. Time (min:sec) elapsed following calcium ionophore addition is shown. Note the prolonged movement of parasites within the PV, probing and distending the PVM (arrows), without egress over the initial 15min.

    Movie s3
    Phase contrast videomicroscopy of ionophore-induced egress by plp1ko tachyzoites from infected fibroblast at 30h post-infection. Time (h:min:sec) elapsed following calcium ionophore addition is shown. Note the prolonged motility within PVs prior to egress, heterogeneity of egress times, and the spherical contraction of several vacuoles.

    Movie s4
    Overlay of fluorescence and phase contrast videomicroscopy of induced egress from a dually infected fibroblast by GFP-positive WT (plp1HA) and non-fluorescent plp1ko tachyzoites at 30h post-infection. Time (h:min:sec) elapsed following calcium ionophore addition is shown.

    Movie s5
    Fluorescence videomicroscopy of induced PVM permeabilization of a WT vacuole expressing DsRed in the PV at 30h post-infection. Time stamp (h:min:sec) indicates time following calcium ionophore addition. Note the rapid loss of fluorescence from the vacuole and diffusion of the signal throughout the host cell cytoplasm.

    Movie s6
    Fluorescence videomicroscopy of induced PVM permeabilization of a plp1ko/PLP1myc strain vacuole expressing DsRed in the PV at 30h post-infection. Time stamp (h:min:sec) indicates time following calcium ionophore addition. Note the rapid loss of fluorescence from the vacuole and diffusion of the signal throughout the host cell similar to WT.

    Movie s7
    Fluorescence videomicroscopy of induced PVM permeabilization of a plp1ko strain vacuole expressing DsRed in the PV at 30h post-infection. Time stamp (h:min:sec) indicates time following calcium ionophore addition. Note that fluorescence remains contained within the PV and only diminishes slowly over time from photobleaching.

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