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. 2011 Nov 3;7(11):e1002356. doi: 10.1371/journal.ppat.1002356

Figure 3. LLO is sufficient to induce the entry of noninvasive L. innocua into HepG2 cells.

Figure 3

(A) HepG2 cells were infected with WT L. monocytogenes (DP10403S, L.m.), L. innocua (L.i.), or L. innocua treated with 1 mM nickel (II) chloride coating buffer in the absence (L.i. buffer) or presence of 5 µM LLO (L.i. buffer + LLO) at MOI = 20 for 30 min at 37°C. Gentamicin was added for 1 h and the intracellular CFUs were enumerated. Results were the mean ± SEM (n≥3) and expressed relative to L. monocytogenes. (B) HepG2 cells were infected with L. innocua (L.i.) or L. innocua phly/prfA* (L.i. phly/prfA*) for 60 min at 37°C (MOI = 100). Gentamicin was added for 30 min and the intracellular CFUs were enumerated. Results were the mean ± SEM (n≥3) and expressed relative to L. i. phly/prfA*. (C) Hemolytic activities of L. monocytogenes (DP10403S) and L. innocua phly/prfA* measured at 37°C for 30 min, pH 7.4. Results were the mean ± SEM (n≥3). (D) Equivalent amounts of L. monocytogenes and L. innocua lysates and proteins precipitated from their culture supernatants were analyzed by western blotting using an anti-LLO primary antibody. A representative experiment (of 3) is presented.