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. 2020 Mar 13;8:153. doi: 10.3389/fcell.2020.00153

FIGURE 4.

FIGURE 4

Identification of RND3 core promoter region and FOXD3 as a transcriptional enhancer for RND3 in HTR-8 cells. (A) Specific primers for sequences upstream of the RND3 transcription initiation site were designed and (B) cloned into pGL3-basic vector. (C) Relative luciferase activity of F1–F5 and basic plasmids. *P < 0.05, ****P < 0.0001. (D,E) ChIP assay of the combination of RND3 promoter. *P < 0.05 vs. the IgG group. (F) Relative luciferase activity of F2–F4 and basic plasmids in HTR-8 cells at 48 h after transfection with control vector or RND3 overexpression vector. *P < 0.05 vs. Vector group. (G) Prediction of two FOXD3 binding sites in the RND3 core promoter region (–663 to –572) (underlined) and construction of the FOXD3 binding site mutant reporters. (H) Relative luciferase activity of RND3 reporter plasmids (WT) and FOXD3 binding site mutant reporters (Mut-1 and Mut-2). Data represent the means ± SD of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 vs. Vector group or FOXD3 group.