Table 1.
Details of the primary antibodies used in this study.
Figure 1.
Heat-map diagram showing gene expression profiles in MS and MS9II cells.
The figure represents data obtained using mouse AD-PCR-Array of 87 selected genes involved in APP/Aβ metabolism, cholesterol metabolism, lysosomal enzyme and cell signalling. Transcriptional levels are colored yellow and different shades of red for significant up-regulations, different shades of green for significant down-regulations and grey for no alteration in MS9II cells compared to MS cells. Of the 87 genes evaluated, 54 transcripts are up-regulated and 9 genes are down-regulated, while remaining 24 genes remained unaltered in MS9II cells compared to MS cells (A). Pie-chart showing percentage of up- and down-regulated genes in MS9II cells compared to MS cells. Gene expression levels are colored yellow and shades of red for significant up-regulation, various shades of green for significant down-regulations and grey for no alteration. As evident from the pie-charts, several genes are differentially altered following overexpression of the human IGF-II receptor in MS9II cells (B). The data included in the heat-map diagram were obtained from four different experiments.
Table 2.
List of selected genes for Mouse Alzheimer’s disease real-time RT-PCR array.
Figure 2.
Transcript and protein expression levels of IGF-II receptor (A, B), IGF-II (C, E) and APP (D, F) in MS and MS9II cells.
Increased levels and expression of IGF-II receptor in MS9II vs MS cells are validated using Western blotting and immunofluorescence staining respectively (A, B). Histograms showing decreased level of Igf2 mRNA (C) and increased level of App mRNA in MS9II cells compared to MS cells as obtained using AD-PCR-Array. Immunoblots and respective histograms validating decreased levels of IGF-II and increased levels of APP in MS9II cells. The protein levels were normalized to the β-actin and the values from four different experiment are expressed as means ± SEM, **p<0.01. Scale bar = 10 µm.
Figure 3.
Transcript and protein expression levels of ADAM9 (A, C), BACE1 (B, D), PS1 (E, G) and APH-1 (F, H) in MS and MS9II cells.
Histograms showing unaltered levels of Adam9 mRNA (A) and increased levels of Bace1 mRNA (B) in MS9II cells compared to MS cells. Immunoblots and respective histograms validating unchanged ADAM9 (C) and increased BACE1 (D) levels in MS9II cells. Histograms showing increased mRNA levels for Psen1 (E) and Aph1a (F) in MS9II cells compared to MS cells. Immunoblots and respective histograms showing unaltered protein levels of PS1 (G) and APH-1 (H) in MS9II cells. The protein levels were normalized to the β-actin and the values from four different experiments are expressed as means ± SEM, *p<0.05, **p<0.01.
Figure 4.
Transcript and protein expression levels of IDE (A, C), PLAU (B, D), GSK-3β (E, G) and CDK5 (F, H) in MS and MS9II cells.
Histograms showing decreased levels of Ide mRNA (A) and unaltered levels of Plau mRNA (B) in MS9II cells compared to MS cells. Immunoblots and respective histograms validating decreased IDE (C) and unchanged uPA (D) levels in MS9II cells. Histograms showing increased mRNA levels for Gsk3b (E) and Cdk5 (F) in MS9II cells compared to MS cells. Immunoblots and respective histograms showing marked increase in protein levels of GSK-3β (G) and CDK5 (H) in MS9II cells. The protein levels were normalized to the β-actin and the values from four different experiments are expressed as means ± SEM, *p<0.05, **p<0.01.
Figure 5.
Transcript and protein expression levels of APOE (A, C), ABCA1 (B, D), LRP1 (E, G) and LRP6 (F, H) in MS and MS9II cells.
Histograms showing increased levels of Apoe mRNA (A) and decreased levels of Abca1 mRNA (B) in MS9II cells compared to MS cells. Immunoblots and respective histograms validating increased APOE (C) and decreased ABCA1 (D) levels in MS9II cells. Histograms showing unaltered levels of Lrp1 mRNA (E) and increased levels of Lrp6 mRNA (F) in MS9II cells compared to MS cells. Immunoblots and respective histograms showing unchanged LRP1 but decreased levels of LRP6 in MS9II cells. The protein levels were normalized to the β-actin and the values from four different experiments are expressed as means ± SEM, *p<0.05, **p<0.01.
Figure 6.
Transcript and protein expression levels of cathepsin B (A, C), cathepsin D (B, D) and β-glucorinidase (E, F) in MS and MS9II cells.
Histograms showing decreased levels of Ctsb mRNA (A) and Ctsd mRNA (B), but increased levels of Gusb mRNA (E) in MS9II cells compared to MS cells. Immunoblots and respective histograms showed decreased levels of pro-cathepsin B and D but increased levels of mature cathepsins B and D in MS9II cells than in MS cells. Immunoblot analysis of β-glucuronidase level, consistent with mRNA, was enhanced in MS9II cells. The protein levels were normalized to the β-actin and the values from four different experiments are expressed as means ± SEM, *p<0.05, **p<0.01.