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. 2006 Jan 3;103(2):443–448. doi: 10.1073/pnas.0503839103

Fig. 3.

Fig. 3.

PPARγ modulates BACE1 promoter activity. (A) Luciferase activities of a 1.5-kb BACE1 promoter/luciferase reporter construct transfected in PPARγ wild-type, knockout, and knockout transfected with PPARγ cDNA MEF. Cells were treated with by IFN-γ (1 ng/ml) + TNF-α (30 ng/ml) with or without ibuprofen (IBU) (10 μM), n = 5. (B) NSAIDs and PPARγ agonists inhibit transcription of BACE1 promoter. N2a-sw cells transfected with BACE1 promoter and incubated with IBU, Pio, Indo, Napro, GW0072X (1 μM), BAY11-7082, and sulindac sulfide n = 4, all at 10 μM concentration. (C) PPARγ transfection inhibits BACE1 promoter activity. N2a-sw cells were transfected with BACE1 promoter construct and PPARγ1, PPARγ2, and PPARγ2 E499Q cDNA and incubated with or without IBU (10 μM); n = 4. (D) MEF transfected with BACE1 promoter control and mutated at the PPRE site were incubated with IBU (10 μM). (E) N2a cells transfected with BACE1 promoter control and mutated at the PPRE site were incubated with IBU (10 μM). Columns represent mean ± SEM, n = 4. Asterisks, significant differences between wild-type cells and treated cells. #, differences between transfected cells treated or untreated. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; #, P ≤ 0.05; ##, P ≤ 0.01, ANOVA followed by a Tukey post hoc test. (F) Gel-shift analysis with the BACE1-PPRE probe using nuclear extract from MEF cells. The major PPARγ-containing complex is indicated by the arrow. Lane 1, MEF wild-type cells; lane 2, MEF PPARγ knockout cells; lane 3, MEF PPARγ knockout cells transfected with PPARγ1 cDNA; lane 4, MEF wild-type cells incubated with a excess of unlabeled BACE1-PPRE probe. (G) Gel shift using HEK293 cells transfected with human PPARγ2 cDNA. Lane 1, control; lane 2, supershift analysis; lane 3, molar excess of unlabeled BACE1-PPRE probe; lane 4, labeled mutant BACE1-PPRE probe (BACE1-PPREM).