Figure S4.
Redistribution of lysosomes during reovirus infection in other cell lines and in HBMECs infected by other reovirus strains. (A–C) HeLa cells and Vero cells were adsorbed with reovirus T1L M1-P208S at MOIs of 1 and 5 PFUs/cell, respectively. At 18 hpi, cells were fixed and processed for confocal microscopy and EM. (A and B) Immunofluorescence images of uninfected and reovirus-infected cells stained with antibodies specific for LAMP-1 (red) and σ3 (rabbit VU219, green). Lysosomes are recruited to VIs (asterisks) in reovirus-infected HeLa cells and Vero cells. Scale bars, 10 µm. (C) EM images showing VIs surrounded by SOs containing virions (arrows). Scale bars, 200 nm. (D–G) HBMECs were adsorbed with reovirus strains T1L and T3D at an MOI of 1 PFU/cell. At 18 hpi, cells were fixed and processed for fluorescence and EM. (D) Confocal images of uninfected and reovirus-infected cells labeled with antibodies specific for LAMP-1 (red) and reovirus σ1 protein (mouse monoclonal antibody 5C6 specific for T1L σ1 and mouse monoclonal antibody 9GB5 specific for T3D σ1, green). Lysosomes reorganize adjacent to VIs (asterisks) during reovirus T1L and T3D infection. Scale bars, 10 µm. (E and F) TEM images showing uninfected and reovirus T1L- and T3D-infected cells. SOs and MCs are observed adjacent to VIs in infected cells. N, nucleus. Scale bars, 500 nm. (G) Quantification of lysosome redistribution around inclusions as detected by confocal microscopy of infected cells.

Redistribution of lysosomes during reovirus infection in other cell lines and in HBMECs infected by other reovirus strains. (A–C) HeLa cells and Vero cells were adsorbed with reovirus T1L M1-P208S at MOIs of 1 and 5 PFUs/cell, respectively. At 18 hpi, cells were fixed and processed for confocal microscopy and EM. (A and B) Immunofluorescence images of uninfected and reovirus-infected cells stained with antibodies specific for LAMP-1 (red) and σ3 (rabbit VU219, green). Lysosomes are recruited to VIs (asterisks) in reovirus-infected HeLa cells and Vero cells. Scale bars, 10 µm. (C) EM images showing VIs surrounded by SOs containing virions (arrows). Scale bars, 200 nm. (D–G) HBMECs were adsorbed with reovirus strains T1L and T3D at an MOI of 1 PFU/cell. At 18 hpi, cells were fixed and processed for fluorescence and EM. (D) Confocal images of uninfected and reovirus-infected cells labeled with antibodies specific for LAMP-1 (red) and reovirus σ1 protein (mouse monoclonal antibody 5C6 specific for T1L σ1 and mouse monoclonal antibody 9GB5 specific for T3D σ1, green). Lysosomes reorganize adjacent to VIs (asterisks) during reovirus T1L and T3D infection. Scale bars, 10 µm. (E and F) TEM images showing uninfected and reovirus T1L- and T3D-infected cells. SOs and MCs are observed adjacent to VIs in infected cells. N, nucleus. Scale bars, 500 nm. (G) Quantification of lysosome redistribution around inclusions as detected by confocal microscopy of infected cells.

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