Fig. 5

NFE2 regulates the TFF1-mediated NF-κB pathway by upregulating miR-423-5p in ARPE-19 cells. a The relative levels of NFE2 were measured by qRT-PCR, which showed that NFE2 was upregulated in DR group. For (b) and (c), ARPE-19 and RPE-J cells were treated with NG or HG. The relative mRNA (b) and protein levels (c) of NFE2 were measured by qRT-PCR and Western blot, respectively. The data showed that expression of NFE2 was significantly increased in the HG group. For (d) and (e), ARPE-19 and RPE-J cells were divided into five groups: control group, si-NC group, si-NEF2 group, p-NC group and p-NEF2 group. d Results from qRT-PCR manifested that si-NEF2 or p-NEF2 was successfully transfected. e miR-423-5p expression were measured by qRT-PCR, which showed that knockdown of NFE2 significantly inhibited miR-423-5p expression, whereas the overexpression of NFE2 significantly promoted miR-423-5p expression. f A dual-luciferase reporter experiment was performed to assess the luciferase activity of reporters in the indicated cells. The relative luciferase activity is the ratio of the luciferase activity in each test cell to that in control cells. g The relative protein levels of TFF1, p-p65, p-IκBα and p-IKKα/β were assessed by Western blot. The results showed that overexpression of NFE2 inhibited the expression of TFF1, and knockdown of miR-423-5p increased the expression of TFF1. The knockdown of TFF1 activated the NF-κB pathway, and the knockdown of miR-423-5p inhibited the NF-κB pathway via TFF1. Overexpressed of NFE2 activates the NF-κB pathway through the miR-423-5p-mediated regulation of TFF1. Data are presented as mean ± SD. Three replicate wells were used per sample in each experiment and experiments were replicated three times. * P < 0.05; ** P < 0.01