Figure 2.

Prx2 is phosphorylated at Thr89 by cdk5 in glutamate model and causes neuronal death. a) Phosphorylation of Prx2 is attenuated in p35 deficient CGNs compared to cultures from wild type (Wt) littermates following glutamate (Glu) excitotoxicity. Cultures were fixed with 4% PFA after glutamate exposure and immunostained for pT89Prx2. b) Quantification of surviving CGNs after glutamate exposure. CGNs from each mouse (Wt n=7; p35−/− n=4) were plated on 3 wells and intensity of fluorescent signal in soma of neurons was measured by image analyses from 60 neurons in 4 random fields/well. c) AV-mediated expression of WtPrx2, Prx2T89A, Prx2T89E in CGNs was shown by Western blot analysis using anti-Flag antibody. d) Overexpression of Prx2 protects CGNs against glutamate. 5-day plated CGNs were infected with AV expressing Wt, phosphorylation resistant (T89A) and phospho-mimicking (T89E) mutants of Prx2 or only GFP as control and exposed to glutamate 2 days after infection. Fixed cultures were then counted for live GFP expressing neurons. Graph is representative of n=3 experiments. Data is presented as mean ±SEM (*Student’s t-Test, p<0.05).