Figure 8.

Inhibition of PKD negates HSP27 overexpression-afforded neuroprotection against OGD. Cortical neuron cultures transduced with AAV constructs (A) or derived from transgenic mice (B–D) were exposed to the PKD inhibitor CID755673 for 30 min (A) or transduced with shRNA targeting PKD (PKDt) or a scrambled control (PKDsc) for 3 d (B), and then subjected to OGD. Cell viability (cell live/dead kit) and cell death (LDH release) were assessed 24 h after OGD. A, B, PKD inhibition with CID755673 or PKD knockdown significantly blocked the neuroprotective effects of HSP27, but not of HSP27-D, following OGD. C, Overexpression of human PKD1, but not PKDdn, restored HSP27-afforded neuroprotection against OGD in PKD-knockdown neurons. All quantitative data are mean ± SEM, with n = 9/group from 3 independent experiments. *p < 0.05 and **p < 0.01 compared with OGD cultures either transduced with empty vector (A) or derived from wild-type (nontransgenic) animals (B, C), and #p < 0.05 and ##p < 0.01 as indicated by the horizontal brackets.