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. Author manuscript; available in PMC: 2015 Jul 1.
Published in final edited form as: Cell Microbiol. 2014 Jan 24;16(7):1068–1079. doi: 10.1111/cmi.12260

Figure 4. Listeria impairs host Cdc42 activity.

Figure 4

A. Effect of bacterial infection on Cdc42 or Rac1 activity. Caco-2 BBE1 cells were either left uninfected (-) or infected with wild-type or ΔinlC bacterial strains for 5.5 hours. Cell lysates were used for measurement of Cdc42-GTP (i) or Rac1 (ii) levels using an ELISA-based method. Statistical analysis by ANOVA indicated P = 0.0001. *, P<0.05 relative to uninfected cells. B. Cdc42 activity in control or Tuba-depleted cells infected with wild-type or ΔinlC Listeria strains. Caco-2 BBE1 cells were treated with a non-targeting control siRNA or an siRNA directed against Tuba mRNA. Approximately 72 hours after transfection, cells were infected with the indicated Listeria strains for 5.5 hours and solubilized in G-LISA lysis buffer for determination of Cdc42-GTP levels (i). Statistical analysis by ANOVA indicated P = 0.0001. *, P<0.05 relative to uninfected cells treated with control siRNA (ii). In parallel with these infection experiments, a duplicate plate of transfected cells was solubilized in RipA buffer for confirmation of Tuba depletion. The GTPase activity data in A or B are mean relative Cdc42-GTP or Rac1-GTP amounts +/− SEM from three experiments. Relative Cdc42-GTP or Rac1-GTP values were obtained by normalizing absolute values to those from uninfected cells (A) or uninfected cells treated with control siRNA (B).