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. Author manuscript; available in PMC: 2009 Nov 15.
Published in final edited form as: Cell. 2008 Nov 14;135(4):691–701. doi: 10.1016/j.cell.2008.09.024

Figure 5. Hsp33 Increases HOCl-Resistance of E. coli and V. cholerae strains.

Figure 5

A. E. coli wild type BB7222 (closed circles) and the hslO deletion strain JW176 (open circles) were cultivated in LB medium at 37°C. B. E. coli rpoH deletion strain BB7224 containing an empty pBAD vector and BB7224 overexpressing Hsp33 under pBAD control were cultivated in LB medium containing 0.4% w/v arabinose at 30°C. C. Vibrio cholerae O395 and the corresponding hslO deletion strain were cultivated in LB medium at 37°C. After reaching exponential growth, the individual cultures were washed with phosphate buffer, supplemented with the indicated HOCl concentrations and incubated for 20 min at 30°C. Cell viability was analyzed by preparing serial dilutions of the cultures and spotting them onto LB plates. Inset. In vivo thiol status of Hsp33. BB7222 was treated with the indicated concentrations of HOCl. After 10 min of stress treatment, samples were removed and all in vivo reduced cysteines were modified with iodoacetamide (IAM). After TCA precipitation and DTT reduction, the 490 Da thiol-specific molecule AMS was used to alkylate all in vivo oxidized cysteines. Hsp33 was visualized by western blot analysis.