Ctively). There was no difference between the 0?2 cm and 2? cm areas (p = 0.3).Figure 3. Shows the optical densities for HSP 70 expression in three different placenta zones for all patients. The upper panel shows non-labor (n = 6 patients) and the lower panel shows labor (n = 6 patients). doi:10.1371/journal.pone.0054540.gHSP70 is Upregulated in Labor and PreeclampsiaThe second set of experiments was designed to test whether there was a difference in HSP 70 expression between labor and non-labor groups for each of the three sites. Figure 4 shows representative blots 11967625 of non-labor versus labor for the three different areas of the placenta (upper panel 0? cm, middle panel 2? cm and lower panel 4? cm). Figure 5 shows an interaction plot for HSP 70 showing the Salmon calcitonin relationship between the means of the 3 different areas of the placenta sampled (0?, 2? and 4? cm) and the two patient groups (Non-labor solid line (n = 6 patients); labor broken line (n = 6 patients)). Individual groups were then compared using the student’s t test. HSP 70 was significantly increased in the labor group when compared to the non-labor group at the 2? cm site (p,0.005). There was no significant difference in HSP 70 expression between non-labor and labor at the 0? cm (p = 0.99) or the 4? cm (p = 0.06) sites. The third set of experiments was designed to test the difference between HSP70 expression in normotensive pregnancies and pregnancies complicated by preeclampsia. Sample representative blots are shown in Figure 6 for some of the patients. The data is summarised in Table 2. There was a significant increase in HSP 70 expression in the preeclampsia non-labor group (n = 4 patients) compared to the control non-labor group (n = 6) in the 0? cm site (p = 0.003). This difference was not seen for at the 2? cm site. Next the labor groups were compared. There was no significant difference between the control labor (n = 6) and preeclampsia labor groups (n = 5) at the 0? cm sites (p = 0.31) however there was a significant increase in HSP 70 expression in the control labor group (n = 6) compared with the preeclampsia labor group at the 2? cm site (n = 6) (p = 0.001). The next of experiments (Figure 7) was designed to determine if there was any difference in HSP 70 expression in second versus third trimester preeclampsia cases. For all cases combined there were no significant differences noted for either the 0? cm sites (median optical density second trimester 24.8), (median optical density third trimester 26) (p = 0.47, 95 C.I. ) or the 2? cm sites (median optical density second trimester 19.9), (median optical density third trimester 19.3) (p = 0.72, 95 C.I.). The final Castanospermine experiment was performed to confirm that the scanning densitometry provided similar results to other quantitative methods. To do this confirmatory experiments were performed as follows. The labour group samples used in experiment one were repeated as above however this time the signals were quantified using the BioRad gel documentation ECL imager system, removing the need for autoradiographs. As forFigure 5. Shows an interaction plot for HSP 70 showing the relationship between the means of the 3 different areas of the placenta sampled (0?, 2? and 4? cm) and the two patient groups. Non-labor solid line(n = 6 patients); labor broken line (n = 6 patients). doi:10.1371/journal.pone.0054540.gTable 2. Shows the median optical density for each group of patients and p value for each comparison from all patients combined.Ctively). There was no difference between the 0?2 cm and 2? cm areas (p = 0.3).Figure 3. Shows the optical densities for HSP 70 expression in three different placenta zones for all patients. The upper panel shows non-labor (n = 6 patients) and the lower panel shows labor (n = 6 patients). doi:10.1371/journal.pone.0054540.gHSP70 is Upregulated in Labor and PreeclampsiaThe second set of experiments was designed to test whether there was a difference in HSP 70 expression between labor and non-labor groups for each of the three sites. Figure 4 shows representative blots 11967625 of non-labor versus labor for the three different areas of the placenta (upper panel 0? cm, middle panel 2? cm and lower panel 4? cm). Figure 5 shows an interaction plot for HSP 70 showing the relationship between the means of the 3 different areas of the placenta sampled (0?, 2? and 4? cm) and the two patient groups (Non-labor solid line (n = 6 patients); labor broken line (n = 6 patients)). Individual groups were then compared using the student’s t test. HSP 70 was significantly increased in the labor group when compared to the non-labor group at the 2? cm site (p,0.005). There was no significant difference in HSP 70 expression between non-labor and labor at the 0? cm (p = 0.99) or the 4? cm (p = 0.06) sites. The third set of experiments was designed to test the difference between HSP70 expression in normotensive pregnancies and pregnancies complicated by preeclampsia. Sample representative blots are shown in Figure 6 for some of the patients. The data is summarised in Table 2. There was a significant increase in HSP 70 expression in the preeclampsia non-labor group (n = 4 patients) compared to the control non-labor group (n = 6) in the 0? cm site (p = 0.003). This difference was not seen for at the 2? cm site. Next the labor groups were compared. There was no significant difference between the control labor (n = 6) and preeclampsia labor groups (n = 5) at the 0? cm sites (p = 0.31) however there was a significant increase in HSP 70 expression in the control labor group (n = 6) compared with the preeclampsia labor group at the 2? cm site (n = 6) (p = 0.001). The next of experiments (Figure 7) was designed to determine if there was any difference in HSP 70 expression in second versus third trimester preeclampsia cases. For all cases combined there were no significant differences noted for either the 0? cm sites (median optical density second trimester 24.8), (median optical density third trimester 26) (p = 0.47, 95 C.I. ) or the 2? cm sites (median optical density second trimester 19.9), (median optical density third trimester 19.3) (p = 0.72, 95 C.I.). The final experiment was performed to confirm that the scanning densitometry provided similar results to other quantitative methods. To do this confirmatory experiments were performed as follows. The labour group samples used in experiment one were repeated as above however this time the signals were quantified using the BioRad gel documentation ECL imager system, removing the need for autoradiographs. As forFigure 5. Shows an interaction plot for HSP 70 showing the relationship between the means of the 3 different areas of the placenta sampled (0?, 2? and 4? cm) and the two patient groups. Non-labor solid line(n = 6 patients); labor broken line (n = 6 patients). doi:10.1371/journal.pone.0054540.gTable 2. Shows the median optical density for each group of patients and p value for each comparison from all patients combined.
