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Figure 3 Escape into the cytosol and subsequent intracellular replication during an infection of J774 macrophages with Listeria monocytogenes ΔactA (diamonds),

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Presentation on theme: "Figure 3 Escape into the cytosol and subsequent intracellular replication during an infection of J774 macrophages with Listeria monocytogenes ΔactA (diamonds),"— Presentation transcript:

1 Figure 3 Escape into the cytosol and subsequent intracellular replication during an infection of J774 macrophages with Listeria monocytogenes ΔactA (diamonds), L. monocytogenes Δhly (triangles), L. innocua P<sub>actA</sub>-gfp prfA hly (squares) and L. innocua P<sub>actA</sub>-gfp prfA (crosses). Cells were infected for 1 h with an MOI of 0.5 bacteria/cell. P<sub>actA</sub> listerial actA gene promoter From: Inefficient Replication of Listeria innocua in the Cytosol of Mammalian Cells J Infect Dis. 2004;189(3): doi: /381206 J Infect Dis | © 2004 by the Infectious Diseases Society of America

2 Table 1 Strains used in a study of replication of Listeria in the cytosol of mammalian cells
From: Inefficient Replication of Listeria innocua in the Cytosol of Mammalian Cells J Infect Dis. 2004;189(3): doi: /381206 J Infect Dis | © 2004 by the Infectious Diseases Society of America

3 Figure 1 Construction of the Listeria–Escherichia coli shuttle vector pJOE carrying the gfp-mut2 gene [22], which is under the control of the listerial actA gene promoter (P<sub>actA</sub>) [16], and the hexose-phosphate transporter gene (hpt) which is under the control of its own promoter [15, 23] From: Inefficient Replication of Listeria innocua in the Cytosol of Mammalian Cells J Infect Dis. 2004;189(3): doi: /381206 J Infect Dis | © 2004 by the Infectious Diseases Society of America

4 Figure 2 Intracellular replication during infection of J774 macrophages with Listeria monocytogenes ΔactA P<sub>actA</sub>-gfp (A) and L. innocua P<sub>actA</sub>-gfp prfA hly (B) at 30 min and 1, 6, and 24 h after infection. Although L. monocytogenes ΔactA P<sub>actA</sub>-gfp showed >5 rounds of replication after 6 h, L. innocua P<sub>actA</sub>-gfp prfA hly replicated a maximum of only 3 times. P<sub>actA</sub> listerial actA gene promoter From: Inefficient Replication of Listeria innocua in the Cytosol of Mammalian Cells J Infect Dis. 2004;189(3): doi: /381206 J Infect Dis | © 2004 by the Infectious Diseases Society of America

5 Figure 4 No. of infected J774 macrophages and mean no
Figure 4 No. of infected J774 macrophages and mean no. of bacteria (Listeria monocytogenes P<sub>actA</sub>-gfp [black bars] and L. innocua P<sub>actA</sub>-gfp prfA hly [white bars]) per cell at different time points after infection. After 6 h, a majority of L. monocytogenes–infected cells contained ⩾16 bacteria/cell, whereas the no. of bacteria in L. innocua–infected cells was never >6–8 bacteria/cell. P<sub>actA</sub> listerial actA gene promoter From: Inefficient Replication of Listeria innocua in the Cytosol of Mammalian Cells J Infect Dis. 2004;189(3): doi: /381206 J Infect Dis | © 2004 by the Infectious Diseases Society of America

6 Figure 5 Growth of Listeria innocua P<sub>actA</sub>-gfp prfA hly hpt (A) and L. innocua P<sub>actA</sub>-gfp prfA hly (B) in minimal medium with glucose (diamonds) with glucose-1-phosphate (squares) or without a carbon source (circles). P<sub>actA</sub> listerial actA gene promoter From: Inefficient Replication of Listeria innocua in the Cytosol of Mammalian Cells J Infect Dis. 2004;189(3): doi: /381206 J Infect Dis | © 2004 by the Infectious Diseases Society of America

7 Figure 6 Adhesion to, internalization by, and intracellular replication in HEp-2 cells after infection with Listeria monocytogenes ΔactA P<sub>actA</sub>-gfp (diamonds), L. innocua P<sub>actA</sub>-gfp prfA hly (squares), L. innocua P<sub>actA</sub>-gfp prfA hly hpt (triangles) and L. monocytogenes Δhpt P<sub>actA</sub>-gfp (crosses). The duration of infection was 1 h, with an MOI of 10 bacteria/cell. Time point 0 (0 h after infection) shows increased adhesion of the hpt-supplemented L. innocua strain From: Inefficient Replication of Listeria innocua in the Cytosol of Mammalian Cells J Infect Dis. 2004;189(3): doi: /381206 J Infect Dis | © 2004 by the Infectious Diseases Society of America

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