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. 2007 Feb 28;35(6):1822–1832. doi: 10.1093/nar/gkm060

Figure 3.

Figure 3.

Effects of the hilE/mlc/double mutation on Salmonella invasiveness and hilD expression. (A) mRNA level of hilE gene determined by real-time PCR analysis. SL1344 (wt) and SR1304 (mlc−) were grown in LB medium without shaking. Total RNA was isolated from aliquots of cells obtained at the exponential phase. Expression levels of the target genes were normalized to that of 16S rRNA gene. Real-time PCR assay was performed three times in duplicate. (B) HEp-2 cells were infected with SL1344 (wild-type), SR1304 (mlc− mutant), BJ2462 (hilE− mutant) and SR1305 (hilE−/mlc− double mutant), which were grown to exponential phase without shaking. The CFU of intracellular bacteria were assessed 2 h after infection. The values are presented as percentages of the wild-type invasion, which was set at 100%. This assay was performed at least twice in triplicate, to allow calculation of means and standard deviations. (C) Thirty micrograms of total RNA from the indicated strains, extracted at the exponential growth phase in LB medium, were subjected to primer extension analysis.