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. Author manuscript; available in PMC: 2011 Jun 1.

Published in final edited form as: Mol Microbiol. 2010 May 4;76(5):1111–1126. doi: 10.1111/j.1365-2958.2010.07192.x

Role of DegP in LacNAc-BSA-induced loss of bundlin. A) Temporal expression of Bundlin in EPEC E2348/69 (WT), ALN188 (degP::kn) and ALN188 complemented in trans with degP (pCAdegP). The amount of bundlin was determined by densitometric analysis of three independent immunoblots. The data represent average % bundlin, relative to the MBP band density. B) Early LA of E2348/69, ALN188 and ALN188 (pCAdegp) to HEp-2 cells. The bars represent the mean number of Hep-2 cells with LA organisms from three experiments, with error bars representing the standard deviation from the means. No significant difference was seen between the LA of the three strains (p > 0.05) C) Effect of 0.8 mg/mL LacNAc-BSA (L) relative to underivitized BSA (B) on the amount of bundlin present in E2348/69 (WT), ALN188 (degP::kn) and ALN188 complemented with degP in trans (pCAdegP) following 30 minutes of incubation. The blot is representative of three independent experiments. The graph represents the cumulative results from the three experiments, and represents mean bundlin immunoblot band density, relative to MBP band density for each lane of the blots. The error bars represent standard deviations from the means. D) Effect of 0.8 mg/mL LacNAc-BSA, relative to underivitized BSA, on the autoaggregation of E2348/69 (WT), ALN188, ALN188 (pCAdegP) UMD916 (ΔbfpF), or UMD916 (pMSD217). The bars represent the mean autoaggregation indexes for three independent experiments, with error bars representing the standard deviations from the means. **p < 0.001 relative to underivatized BSA. E) Enzymatic activity of recombinant DegP on purified bundlin and lysozyme in the presence or absence of 5mM DTT. Samples were visualized by Coomassie brilliant blue staining of SDS-PAGE gels.

Role of DegP in LacNAc-BSA-induced loss of bundlin. A) Temporal expression of Bundlin in EPEC E2348/69 (WT), ALN188 (degP::kn) and ALN188 complemented in trans with degP (pCAdegP). The amount of bundlin was determined by densitometric analysis of three independent immunoblots. The data represent average % bundlin, relative to the MBP band density. B) Early LA of E2348/69, ALN188 and ALN188 (pCAdegp) to HEp-2 cells. The bars represent the mean number of Hep-2 cells with LA organisms from three experiments, with error bars representing the standard deviation from the means. No significant difference was seen between the LA of the three strains (p > 0.05) C) Effect of 0.8 mg/mL LacNAc-BSA (L) relative to underivitized BSA (B) on the amount of bundlin present in E2348/69 (WT), ALN188 (degP::kn) and ALN188 complemented with degP in trans (pCAdegP) following 30 minutes of incubation. The blot is representative of three independent experiments. The graph represents the cumulative results from the three experiments, and represents mean bundlin immunoblot band density, relative to MBP band density for each lane of the blots. The error bars represent standard deviations from the means. D) Effect of 0.8 mg/mL LacNAc-BSA, relative to underivitized BSA, on the autoaggregation of E2348/69 (WT), ALN188, ALN188 (pCAdegP) UMD916 (ΔbfpF), or UMD916 (pMSD217). The bars represent the mean autoaggregation indexes for three independent experiments, with error bars representing the standard deviations from the means. **p < 0.001 relative to underivatized BSA. E) Enzymatic activity of recombinant DegP on purified bundlin and lysozyme in the presence or absence of 5mM DTT. Samples were visualized by Coomassie brilliant blue staining of SDS-PAGE gels.