To rule out the residual expression of the proteins fromin vitrobacterial growth, we investigated infected mice for longer periods after infection. BALB/c

mice were intraperitoneally infected with 1 × 105CFU bacteria and tissues were collected at 5 days postinfection, prior to the onset of severe diseases associated with infection. Similar results were observed as described above for 18 hours postinfection, which showed that click here proteins PrgI, SopE2, SipB, and SipA were expressed inSalmonellaisolated from both the spleen and cecum, and that SpaO and SptP were preferentially expressed bySalmonellarecovered from the cecum and spleen, respectively (Figure7). Indeed, the expression of the SpaO protein was more than 40 fold higher than that of SptP inSalmonellaisolated from the cecum, while the SptP protein was expressed at least 70 times more than SpaO inSalmonellaisolated from the spleen (Figure7). The protein levels of SopE2 and SipB at 5 days postinfection were higher than those at 18 hours postinfection inSalmonellaisolated from both the cecum and spleen. In contrast,

the levels of PrgI and SipA at 5 days postinfection were lower than those at 18 hours postinfection. It is interesting to note that the expression of SpaO inSalmonellacolonizing the cecum and SptP inSalmonellacolonizing the spleen decreased with the duration of the infection (Figure7). In vivoexpression of Talazoparib datasheet the tagged SPI-1 proteins during oral infection To study whether the tagged SPI-1 proteins are expressed duringSalmonellainfection acquired by the natural route, BALB/c mice

were infected intragastrically with 1 × 105CFU bacteria. Spleens and cecums were collected and the bacteria were recovered at day 7 postinfection, prior to the onset of severe diseases associated with infection. Similar to what was observed inSalmonellafrom intraperitoneally infected mice, PrgI, SopE2, SipB, and SipA were detected inSalmonellaisolated from both the spleen and cecum, while SpaO and SptP were found to be expressed preferentially inSalmonellaisolated from the cecum and spleen, respectively (Figure8). Protein level of DnaK did not appear to be significantly different in bacteria recovered from the spleen and cecum SPTLC1 (data not shown). These results provide direct evidence that PrgI and SipB are expressedin vivoin intragastrically-infected mice. Furthermore, these results suggest that the SpaO and SptP proteins are expressed preferably inSalmonellacolonizing the cecum and spleen respectively during oral infection of mice. Figure 8 Level of the tagged proteins from the internalized bacterial strains T-prgI, T-sipA, T-sptP, T-spaO, T-sopE2, and T-sipB recovered from the spleen and cecum. BALB/c mice were intragastrically infected with 1 × 105CFU of the tagged strains, and internalized bacteria were recovered from the spleen and cecum at 7 days post inoculation.