Fig. 5

Silencing of HSP70 abolished the cytoprotective effect of 17AAG against sevoflurane-induced apoptosis and ROS generation in H4 cells. H4 cells were transfected with vehicle (CON, control), or siRNA against human HSPA1A (HSPA1A siRNA), or non-targeting siRNA (control siRNA), and received treatments as in Fig. 2a–c The expression of HSP70 protein (a, b) and HSPA1A mRNA c in H4 cells was examined by Western blot and RT-qPCR, respectively. d, e Treated cells were stained with Annexin V-FITC and propidium iodide (PI) and analyzed for apoptosis by flow cytometry (lower right quadrant, early apoptosis; upper right quadrant, late apoptosis). (f and g) The generation of ROS in H4 cells was analyzed by flow cytometry as in Fig. 2. Shown representative images of 3 independent experiments (a, d, g). n = 6. Mean ± SD, **P < 0.01 vs. SEV, ## P < 0.01 vs. SEV + 17AAG by one-way ANOVA followed by Bonferroni multiple comparisons test