Fig. 2.

Protein assay of LKB1/AMPK axis: phosphorylated (Active) form of LKB1 (A) and AMPK (B) and the total protein of each (A, B) were analysed via Western blot of the cortical and hippocampus regions of the control, ischaemic only and ischaemia followed by short-term 60 min and long-term recovery of 4 weeks following 2 min CA (n = 6 each). I-2m: 2 min ischaemia, I-2m/R- 60 m: 2 min ischaemia followed by 60 min reperfusion, I-2m/R- 4w: 2 min ischaemia followed by 4 weeks recovery period (One Way ANOVA, *p < 0.05 for p-LKB1 in I-2m vs control, *p < 0.01, for p-LKB1 in I-2m/R- 4w vs control; *p < 0.01, for p-AMPK in I-2m, I-2m/R- 60 m and I-2m/R- 4w vs control). Error bars depict the SD. All values are expressed as percent change relative to control group and were corrected by the Actin level. C) Phosphorylated AMPK (green) presence in control (Fig. 2C a, b, c) and long-term recovery of 4 weeks (Fig. 2C d, e, f) are visualised by immunofluorescent staining. D) DAPI (blue) shows nuclei staining for all cell types (both neurons and glial cells). Distribution of p-AMPK within the neurons and glial cells are shown by immunofluorescent co-staining with neuronal marker for nuclei (NeuN (a), red) or cytoplasmic glial fibrillary acidic protein (GFAP (b), red) from the brain samples of long-term recovery group (2 min ischaemia followed by 4 weeks). Accumulation of p-LKB1 is shown via DAB staining in long-term recovery group (d) compared with control group (c). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)