FIG. 2.

Functional deletion analysis of the human BACE1 gene promoter. (A and D) Schematic diagrams of the BACE1 promoter deletion constructs consisting of a 5′ flanking region with serial deletions cloned into the promoter-less vector plasmid pGL3-basic in front of a reporter gene, the luciferase gene (Luc). Arrow, direction of transcription. The numbers represent the end points of each construct. (B and E) The deletion plasmids shown in panels C and F, respectively, were confirmed by sequencing and restriction enzyme digestion checking, and the digested samples were analyzed on a 1.1% agarose gel. Vector size is 4.7 kb, and the BACE1 gene 5′ flanking fragment insert sizes range from 0.4 to 2.2 kb. (C and F). The constructed plasmids were cotransfected into HEK293T cells with pCH110. Luciferase activity was measured at 48 h by a luminometer. β-Gal activity was used to normalize transfection efficiency. The values represent means ± standard errors of the means (n = 3 to 6). *, P < 0.001 by ANOVA with the post hoc Newmann-Keuls test.