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. Author manuscript; available in PMC: 2015 Nov 3.
Published in final edited form as: Curr Alzheimer Res. 2015;12(9):847–859. doi: 10.2174/156720501209151019105834

Figure 10. (A) USF proteins positively regulate APP promoter activity through an E-box element in the APP promoter.

Figure 10

The E-box element, CAGCTGA (consensus CANNTG) at position −49 base pairs upstream of the transcription start site was mutated to CGTGCTAC by site-directed mutagenesis. Cells were co-transfected with the wild-type APP promoter or APP E-box mutant promoter constructs and USF-1, USF-2 or dominant-negative A-USF expression constructs. Cell homogenates were assayed for fold-change in APP promoter activity as described in Methods. Results are the mean ± SD of six determinations. Statistical significance was determined using the Student’s t test, p < 0.05. (B) ABCA2 overexpression increases mass of USF-1 and USF-2 transcription factors. Protein lysates from N2a, A2 cells were fractionated on 4–12% NuPAGE gels transferred to nitrocellulose and probed by Western blot with the USF-1 antibody (43 kDa) and USF-2 antibody (44 kDa). The results are the mean % change in protein mass in A2 cells compared to N2a cells ± SD ≥ 3 determinations. Statistical significance was determined using the Student’s t test, p < 0.05.