Figure 2.

Autophagosomes accumulate in the cortex from 1.5 h to 1 d after HI. (a) Western blots of PIK3C3/VPS34, Beclin-1, ATG12-ATG5 conjugate, and phospho-ULK1 in cortical tissue lysates from sham-operated and HI piglets at the indicated time points. Each lane corresponds to an individual animal (two per time point). (b–e) Densitometric analysis of (b) PIK3C3/VPS34, (c) Beclin-1, (d) ATG12-ATG5 conjugate, and (e) phospho-ULK1 data from (A) normalized to the loading control, β-actin (n=4, *P<0.05, **P<0.01 versus the sham-operated group). (f–g). Quantitative PCR results of Atg12 mRNA (F) and Beclin-1 mRNA (G). (h) Images of piglet cortical brain sections stained with antibodies against the autophagy marker Beclin-1 and the neuronal marker NeuN. Scale bar=50 μm. (i) Quantification of NeuN-positive cells and Beclin-1 and NeuN double-positive cells normalized to the total cell number (**P<0.01 versus the sham-operated group). The percentages of double-positive versus single-positive cells are indicated at 6 h and 1 d after HI. Data are presented as mean±S.D. (n=3; at least 1000 cells were quantified per piglet per experiment)