Figure 3. LncRNA WT1-AS inhibited miR-375 expression by regulating the transcription factor WT1. (A) Differential miRNA expression heatmap of GSE16759; abscissa, sample number; ordinate, miRNA name; and left tree, miRNA expression level clustering. Each small square in the figure represents the expression of one miRNA in a sample, and the histogram at the top right is the color scale. (B) Prediction of regulatory miRNAs downstream of WT1. The middle part indicates the intersection of predicted WT1 target miRNAs and differentially expressed miRNAs in GSE16759. (C) The differential expression of miR-375 in GSE16759. (D) The expression of miR-375 in Aβ25-35treated SH-SY5Y cells detected by qRT-PCR. (E) Prediction of WT1 binding site in the miR-375 promoter region by combining NCBI (https://www.ncbi.nlm.nih.gov/) and JASPAR (http://jaspar.genereg.net/). (F) After mutation of WT1 in the promoter region of miR-375, dual luciferase reporter assay was used to detect WT1 targeted binding to the promoter region of miR-375 in HEK-293T cells. (G) Detection of WT1 targeted binding to the promoter region of miR-375 by ChIP assay. (H–I) The expression of miR-375 detected by qRT-PCR. *P<0.05; the experimental results are expressed as the mean ± standard deviation. Differences among multiple groups were analyzed using one-way ANOVA followed by Tukey’s post hoc test. The experiment was repeated three times.