Fig. 4: The region 1 kb distal to the APOE 3′UTR acts as a transcriptional repressor and Aβ and SPI1 treatment impacts APOE-associated transcript expression in HMC3 cells.
From: A common 19 bp APOE enhancer deletion is protective against Alzheimer’s disease in African Americans
a Schematic of the luciferase experimental design and constructs. The APOE 3′UTR (APOE 3′UTR only) plus ~400 bp of downstream DNA containing the 19 bp deletion region and SPI1 binding site were cloned into a psiCheck2.2 dual luciferase reporter construct. b Renilla:Firefly luciferase expression data in HMC3 cells after delivery of APOE SPI1 WT sequence (p = 0.0016 relative to the APOE 3′UTR only) or delivery of the 19 bp deletion [p = 0.8252; one-way ANOVA, n = 12 samples/group (3 biological replicates, 4 technical replicates), ± SD]. c–e HMC3 gene expression following SPI1 overexpression. c Effect of SPI1 overexpression on c APOE expression (p = 0.3831, F = 1.789, unpaired two-tailed t-test, n = 12 control and 12 SPI1 biological replicates derived from the average of 4 technical replicates/sample, ± SEM), d APOC1 expression (p = <0.0001, F = 2.987, unpaired two-tailed t-test, n = 11 control and 12 SPI1 biological replicates derived from the average of 4 technical replicates/sample, ± SEM), and e lncRNA ENSG00000280087 expression (p = 0.0170, F = 5.414, unpaired two-tailed t-test, n = 12 control and 10 SPI1 biological replicates derived from the average of 4 technical replicates/sample, ± SEM). f–h HMC3 gene expression following Aβ 1:42 treatment. f Effect of Aβ 1:42 treatment on APOE expression (p = 0.2098, F = 1.243, unpaired two-tailed t-test, n = 12 control and 12 Aβ biological replicates derived from the average of 4 technical replicates/sample, ± SEM), g APOC1 expression (p = 0.0015, F = 4.657, unpaired two-tailed t-test, n = 11 control and 12 Aβ biological replicates derived from the average of 4 technical replicates/sample, ± SEM) and h, lncRNA ENSG00000280087 expression (p = 0.0255, F = 2.502, unpaired two-tailed t-test, n = 12 control and 12 Aβ biological replicates derived from the average of 4 technical replicates/sample, ± SEM) in HMC3 cells.
