S100a9 Knockdown Decreases the Memory Impairment and the Neuropathology in Tg2576 Mice, AD Animal Model (original) (raw)
Figure 2
Aβ and CT induced S100a9 expression in BV2 cells.
(A) Immunostaining for S100a9 (green) and CD11b (red) was performed in the cortex and CA3 of Tg and wild-type mouse brains. DAPI (blue) staining indicates the nuclei. Scale bars represent 20 µm. WT and Tg represent wild-type control and transgenic animals, respectively. (B) The mRNA levels of S100a9 were checked at 24 h, 48 h, and 72 h post-transfection with CT50 or CT99 by RT-PCR. (C) Human S100a9 promoter in pGL3 vector was cotransfected with CT, wtAPP or sweAPP in pcDNA vector and fe65 into SHSY5Y cells. 48 h after transfection, luciferase reporter assays were performed for CT50, CT99, wtAPP and sweAPP, the four S100a9 targets. Luciferase activities were normalized versus the obtained protein concentrations. The histogram shows the ratio of luciferase activity in transfected cells to empty vector-transfected cells (n = 5). (D) RT-PCR of S100a9 induced at 48 h post-treatment with 1 µM and 10 µM CT or 2 µM and 20 µM Aβ. (E) The quantification of RT-PCR normalized to actin (n = 8). (F) Immunocytochemistry; BV2 cells grown on a glass coverslip were treated with CT (5 µM and 10 µM) and Aβ (20 µM) for 48 h. S100a9-positive cells were visualized with FITC (green)-conjugated secondary antibodies. Nuclei were counter-stained with DAPI (blue). Scale bars represent 20 µm. All data are presented as the means±SEM. *P<0.05, **P<0.01 by Student's t –test.