Figure 7.

Srsf6 Was Identified as the Direct Target of miR-146a

(A) 293T cells were transfected with miR-146a mimics, scrambled miRNA (NC), or inhibitor for 24 h, and the relative levels of miR-146a were detected by qRT-PCR. (B) Then the protein levels of Psmd3, Srsf6, Stx3, and Rgs11 in 293T cells were also detected by western blotting analysis; β-actin was used as the control. (C) SH-SY5Y cells were transfected with miR-146a mimics, scrambled miRNA, or inhibitor for 24 h, and the relative levels of miR-146a were detected by qRT-PCR. (D) Then the protein levels of Psmd3, Srsf6, Stx3, and Rgs11 in SH-SY5Y cells were also detected by western blot analysis; β-actin was used as the control. (E) The protein levels of Psmd3, Srsf6, Stx3, and Rgs11 in hippocampi of AD, ADI, and control groups were detected by western blotting analysis; β-actin was used as the control (n = 4). (F–J) Luciferase reporter assays were used to detect the interaction between the putative Psmd3 (F), Stx3 (G), Rgs11 (H), Srsf6-1 (I), and Srsf6-2 (J) binding sites with miR-146a. Relative activity of luciferase constructs bearing either wild-type (wt) or mutant (mut) Psmd3, Srsf6-1, Srsf6-2, or Stx3, Rgs11 3′ UTRs in cells co-transfected with miR-146a mimic is presented as Renilla/firefly luminescence units (RLUs). All data are presented as the mean ± SEM. At least three independent experiments were performed. *p < 0.05; **p < 0.01; ***p < 0.001.