Fig. 1
The expression of EAAC1 in CoCl2-treated SH-SY5Y cells. a SH-SY5Y cells were treated with different concentrations of CoCl2 (0, 50, 100, 150, 200, 300 and 500 µM) for 24 h, which resulted in dose-dependent increases in EAAC1 expression. b Quantitative analysis of EAAC1 immunoreactivity in a. The results are presented as the means ± S.E.M.; n = 8; ***P < 0.001. c Western blotting demonstrated that 100 µM CoCl2 affected EAAC1 protein expression levels. EAAC1 protein expression was significantly increased by 100 µM CoCl2 in a time-dependent manner. d Quantification of the data in c. The densitometry values are shown as ratios relative to the values of the control group (n = 6; *P < 0.05, ***P < 0.001). e The degree of cell death was assessed 36 h after EAAC1-myc transfection using LDH activity in the medium (Mock: n = 6; ***P < 0.001 versus the control group; EAAC1-myc: n = 6; ###P < 0.001 versus the control group; CoCl2 group: ++P < 0.01 versus the Mock group). Representative immunoblots for Myc-tag are shown for transfection with EAAC1-myc. f Illustration of procedure for CoCl2 micro-injections (1 µl-25 mM) into the ventral hippocampus (coordinates from bregma: anterior/posterior − 3.3 mm, medial/lateral + 2.8 mm, dorsal/ventral − 4.0 mm). g Coronal sections of the ventral hippocampus of Saline and CoCl2 groups were stained with anti-EAAC1. Photomicrographs reveal EAAC1 expression in the DG (g), CA1 (i), and CA3 (k) regions of the ventral hippocampus in mice treated with saline (top panels) and CoCl2 (bottom panels). Scale bar, 100 µm; inset, enlarged areas. Scale bar, 25 µm. h Quantification analysis of EAAC1 immunoreactivity in g. j Quantification analysis of EAAC1 immunoreactivity in i. l Quantification analysis of EAAC1 immunoreactivity in k. Data on EAAC1 staining quantification are expressed as the means ± S.E.M.; n = 8; ***P < 0.001, *P < 0.05