Figure 1

Ten percent HS down-regulated NKCC1 expression in peri-ischemic brain tissue. Sample was extracted from the peri-ischemic brain tissue outlined by the green line in (A). NKCC1 mRNA and protein expression in the peri-ischemic brain tissue of ischemic and ischemic + 10% HS rats at 12 h following MCAO and the corresponding control rats (sham-operated). Section (B) shows the graphical representation of the fold changes in NKCC1 mRNA in each group as quantified by normalization to the β-actin as an internal control (n = 6 per group). Section (C) shows NKCC1 (170 kDa) and GAPDH (37 kDa) immunoreactive bands, respectively. (D) This is a bar graph showing significant changes in the relative optical density of NKCC1 to GAPDH in each group. Significant differences in mRNA and protein levels in the peri-ischemic brain tissue of ischemic 12 h, ischemic 12 h + 10% HS are evident when compared with sham-operated ( # P <0.05,* P <0.05, n = 6 per group). Confocal images showing the distribution of GFAP labeled (E, H, K, red), and NKCC1 (F, I, L, green) immunoreactive astrocytes (arrows) in the peri-ischemic brain tissue of ischemic and ischemic + 10% HS rats at 24 h following MCAO and the corresponding control rats (sham-operated). GFAP labeling clearly overlaps NKCC1 immunofluorescence in (G, J and M). Note that NKCC1 expression in the perivascular astrocytes (arrows) is markedly enhanced following MCAO. After treatment with 10% HS, NKCC1 expression is noticeably attenuated. Scale bars: (E-M), 50 μm. GFAP, glial fibrillary acidic protein; HS, hypertonic saline; NKCC1, Na-K-Cl Cotransporter 1; MCAO, middle cerebral artery occlusion.