p53 Inhibition Protects against Neuronal Ischemia/Reperfusion Injury by the p53/PRAS40/mTOR Pathway

Figure 7

Neuronal autophagy was exacerbated by PRAS40 knockdown both in vivo and in vitro. (a) As shown by immunostaining (×100) and TEM in ischemic mouse brains 48 h after MCAO, more LC3 II-positive cells (immunostaining) and autophagosomes (TEM; white arrow) were observed in p53^+/+ WT mice than in p53^+/- or p53^-/- mice. (b) In the ischemic brain 48 h after MCAO, the level of LC3 II was significantly higher in WT mice than in p53^+/- or p53^-/- mice. vs. the WT+MCAO group; ^& vs. the WT+sham group. (c) As shown by immunostaining (×400) of p53^-/- mouse brains 48 h after MCAO, the level of LC3 II and the ratio of LC3 II-positive to RFP-positive cells were much higher in cells pretransfected with PRAS40 shRNA than in those pretransfected with or without scramble shRNA. vs. the p53^-/-+PRAS40 shRNA group. (d) In p53^-/- mouse brains 48 h after MCAO, PRAS40 shRNA administration increased the level of LC3 II much more than that in the control groups treated with or without scramble shRNA. ^# vs. the p53^-/-+PRAS40 shRNA group. –8/group. (e) As shown by immunostaining (×400), the expression of LC3 II in primary cultured p53^-/- mouse neurons were markedly higher when cells were pretransfected with PRAS40 shRNA than when they were pretransfected with or without scramble shRNA.

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