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. 2019 Jun 17;8:e45957. doi: 10.7554/eLife.45957

Figure 2. Cas9+ CIMs as an in vitro model for Listeria monocytogenes infection.

Figure 2.

(a) BMMs (left panel) or CIMs (right) were infected with WT Listeria monocytogenes at MOI = 0.25 and monolayers were visualized with Diff-Quick stain at 8 hr post-infection. Data are representative of two independent experiments. (b) BMMs (top three panels) or CIMs (lower panels) were infected for 5 hr with three strains of L. monocytogenes: WT, Δhly, and ΔactA at MOI = 1.5. Nuclei shown in blue, bacterial cells in red, and F-actin in green. (c) BMMs (left panel) or CIMs transduced with a scramble gRNA were infected with three strains of L. monocytogenes: WT, an autophagy-sensitive strain that lacks ActA, PlcA and PlcB (Mitchell et al., 2018) and Δhly at MOI = 0.25. Bacterial densities were enumerated by CFU at t = 0.5 h and indicated hours post-infection. Data are representative of two independent experiments each performed in triplicate, mean ± SD are shown. (d) CIMs transduced with the indicated gRNAs were infected with WT L. monocytogenes or the autophagy-sensitive L. monocytogenes mutant at MOI = 0.75 and bacterial density was enumerated by CFU at t = 8 hr. Data are representative of two independent experiments each performed in triplicate, mean ± SD are shown.