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. 2019 Nov 28;13:1264. doi: 10.3389/fnins.2019.01264

FIGURE 6.

FIGURE 6

Plek and Sdcbp2 might be the targets of miR-409-5p. (A) The wild-type (WT) and mutated plek or Sdcbp2 binding sites with miR-409-5p. (B) Relative Renilla/luciferase luminescence of a psiCHECK2 vector construct harboring Plek or mutant Plek cotransfected with miR-409-5p in the HEK 293T cells, with empty psiCHECK2 vector as control. (C) Relative Renilla/luciferase luminescence of a psiCHECK2 vector construct harboring Sdcbp2 or mutant Sdcbp2 cotransfected with miR-409-5p in the HEK 293T cells, with empty psiCHECK2 vector as control. The results were shown as the mean ± SD (∗p < 0.05,∗∗p < 0.01). The experiment was repeated independently for at least three times. (D,E) MiR-409-5p mimic (100 nM) or inhibitor (100 nM) was transfected into Neuro-2a cells. After 24 h, plek and sdcbp2 mRNA levels were assessed by RT-qPCR. (F,G) Western blot analysis of Plek and SDCBP2 protein level in primary cortical neurons transfected with miR-409-5p mimic (100 nM) or inhibitor (100 nM) for 72 h. ImageJ software was used to quantify the gray degree values of Plek and SDCBP2. All results were shown as the mean ± SD (∗p < 0.05). The experiment was repeated independently for three times. Unpaired t-test was used to analyze differences.