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. 2020 May 30;10(16):7083–7099. doi: 10.7150/thno.44744

Figure 5.

Figure 5

OA-induced ANGPTL4 regulates NOX4 transcriptional activity through the AP-1 binding site on the NOX4 promoter. (A) Dual-luciferase reporter assay was performed to analyze the activation of NOX4 promoter (4.7 kb length) in SW480 cells. Cells were treated with 200 µM OA for the indicated period of time (i). The level of remaining NOX4 mRNA was analyzed by real-time quantitative PCR in SW480 cells treated with or without 200 µM OA for 3 h followed by incubation with 4 μM actinomycin D (ActD) for the indicated period of time (ii). (B) Dual-luciferase reporter assay was performed in cells transfected with a series of 5'-truncated NOX4 promoters followed by treatment with 200 µM OA for 16 h. (C-D) Dual-luciferase reporter assay and ELISAs were performed in SW480 cells transfected with the wild-type (WT) NOX4 promoter or with AP-1 binding site mutation (AP-1m) (C), and 20 nM siANGPTL4 (siANG) (D) (i, iii) or expression vector of ANGPTL4 (flANG) (D) (ii, iv), followed by treatment with 200 µM OA for 16 h. Firefly luciferase activity was determined and normalized to Renilla luciferase activity. The data are presented as the mean ± SEM. P-values were determined using a two-tailed Student's t-test. ***P < 0.001 (n=3).