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. 2017 Dec 5;8(67):111882–111901. doi: 10.18632/oncotarget.22948

Figure 5. Analysis of the miR-124 promoter and transcription factor Sp1 binds to miR-124 promoter.

Figure 5

(A) RAW264.7 cells were co-transfected with pRL-TK and a miR-124 promoter reporter plasmid containing various lengths of the miR-124 promoter. The first nucleotide of mature pre-miR-124 is assigned +1. Right panel shows the relative luciferase activities of these plasmids. Luciferase activity of the mock transfected empty vector pGL3-Basic was used for normalization. The data in graph were shown in mean ± SEM of 3 different transfections. (B) Schematic diagram of miR-124 genomic location in mouse chromosome 2. The promoter region from pre-197 to pre-27 was used for analysis of potential transcription factor binding sites. Two Sp1 binding sites are indicated as diamonds. The sequences of mutated sites are shown under the locus diagram. (C) Schematic representation of deletion mutants 1 and 2 in the full-length miR-124 promoter. (D) RAW264.7 cells were co-transfected with the above promoter reporter constructs and pRL-TK. Post-transfection for 36h, samples were collected and analyzed for dual luciferase activity. (E) Real-time qPCR and Western blot analysis of the expression of Sp1 after transfection with ICAM-1 siRNA and negative control. (* p < 0.05, ** p <0.01, *** p <0.001).