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. 2014 Jul;184(7):2066–2081. doi: 10.1016/j.ajpath.2014.03.014

Figure 2.

Figure 2

Expression levels of cholesterol-transporting polypeptides. A: Representative immunoblots of StARD3 and StAR showing an increased presence of these cholesterol carriers in brain samples from APP/PS1 mice compared with 10-month-old WT mice. B: mRNA levels of StARD4 and StARD5 in brain samples from WT and APP/PS1 mice at 10 months of age analyzed by quantitative RT-PCR. Absolute mRNA values were determined, normalized to 18S rRNA, and reported as relative levels compared to the expression in WT mice. C: Representative confocal images of StAR (red) and MAP2 (green) immunofluorescence labeling of hippocampal sections from 10-month-old WT mice and APP/PS1 mice. D: Representative confocal images of hippocampal sections from 10-month-old APP/PS1 mice immunostained with anti-StAR (red) and anti-MAP2 or anti–glial fibrillary acidic protein (GFAP; green). The extent of colocalization was quantified using the linear Pearson correlation coefficient (rp); the resulting scatterplot images are shown on the right. A value of 1.0 indicates complete colocalization of two fluorescent signals. Note that StAR preferentially colocalizes with neurons labeled with anti-MAP2 compared with astrocytes labeled with anti-GFAP. E: Western blot analysis of StARD3 and StAR expression in brain from 10-month-old WT mice and Tg–SREBP-2 mice at the indicated ages. F: Total cholesterol in ER and MAMs from brain extracts of 10-month-old WT and APP/PS1 mice. G: Representative immunoblots showing the expression of calreticulin (calregulin), the sigma-1 receptor (σ1R), phosphofurin acidic cluster sorting protein 2 (PACS-2), and the mitochondrial proteins cyclooxygenase 4 (COX IV), voltage-dependent anion-selective channel protein (VDAC), and mitofusin 2 (MFN2) in ER, MAMs, and mitochondria from 10-month-old WT and APP/PS1 (A/P) mice. In all Western blot analyses the densitometric values of the bands representing the specific protein immunoreactivity were normalized with the values of the corresponding β-actin bands or Ponceau Staining. ∗P < 0.05, ∗∗P < 0.01 versus WT mice values by one-way analysis of variance with the Dunnett post hoc test (A and E) and the unpaired, two-tailed Student's t-test (B, C, and G) (n = 3 to 6). Scale bars: 15 μm (C and D). mito., mitochondria; mth, month.