GW3965 increases the nestin immunoreactivity and the proliferating cells number in hippocampus of 3xTg-AD mice.

(A) Representative micrographs of multiple immunofluorescence of ApoE (Green) plus nestin (Red) and side by side only nestin (Red) immunofluorescence in DG using confocal microscopy for treated and untreated WT and 3xTg-AD mice. (B) Quantification of nestin positive immunofluorescence in the SGZ of DG. (C) Representative micrographs of ApoE (Green) and nestin (Red) immunofluorescence (60X), the colocalization threshold of ApoE and nestin indicates that there is a population of cells with increased ApoE expression. (D) Representative micrographs of DG of treated and untreated WT and 3xTg-AD mice showing the immunofluorescence of ApoE and side by side phospho-Histone H3 (Ser10) (Red) plus Hoechst (Blue). (E) Quantification of the number of positive-nuclei labeled with pHH3 (Red) and Hoechst (Blue). (F) Representative micrographs of ApoE (Green) and phospho-H3 (Red) immunofluorescence and nuclei Hoechst (Blue,60X), showing that there are proliferating cells with increased ApoE expression. Data are expressed as mean ± S.E.M. Statistical analysis was performed by one-way ANOVA followed by Bonferroni post test: *:p<0.05; **: p<0.01, n = 4 per group.

(A) 采用共聚焦显微镜（confocal microscopy）拍摄经处理与未处理的野生型（wild type, WT）及3xTg-阿尔茨海默病（3xTg-AD）小鼠齿状回（dentate gyrus, DG）的代表性显微图像：一组为载脂蛋白E（Apolipoprotein E, ApoE，绿色）联合巢蛋白（nestin，红色）的多重免疫荧光（immunofluorescence）染色，另一组为仅标记巢蛋白（红色）的单重免疫荧光（immunofluorescence）染色，二者并列展示。
(B) 对DG颗粒下层亚区（subgranular zone, SGZ）内巢蛋白阳性免疫荧光信号的定量分析结果。
(C) 载脂蛋白E（绿色）与巢蛋白（红色）免疫荧光染色的代表性显微图像（60倍物镜）；通过载脂蛋白E与巢蛋白的共定位（colocalization）阈值分析，可观察到存在一类载脂蛋白E表达水平升高的细胞群。
(D) 经处理与未处理的野生型及3xTg-AD小鼠齿状回的代表性显微图像，并列展示两组染色结果：一组为载脂蛋白E免疫荧光染色，另一组为同时标记磷酸化组蛋白H3（Ser10）（phospho-Histone H3 (Ser10)，红色）与Hoechst（蓝色）的免疫荧光染色。
(E) 对磷酸化组蛋白H3（红色）与Hoechst（蓝色）双标记的阳性细胞核数量的定量分析。
(F) 载脂蛋白E（绿色）、磷酸化组蛋白H3（红色）与细胞核染料Hoechst（蓝色）的免疫荧光染色代表性显微图像（60倍物镜），结果显示存在载脂蛋白E表达水平升高的增殖细胞群。
数据以平均值±标准误（standard error of the mean, S.E.M.）表示，统计分析采用单因素方差分析（one-way ANOVA）结合Bonferroni事后检验：*:p<0.05; **: p<0.01，每组n=4。