Research Paper Volume 12, Issue 20 pp 20728—20742

Figure 6. miR-338-5p ameliorated neuronal apoptosis by directly targeting BCL2L11. (A) TargetScan was used to predict the binding sites of miR-338-5p within the 3ʹ-UTR of BCL2L11. (B) Overexpressing miR-338-5p resulted in a remarkable decrease in luciferase activity of BCL2L11-WT and exerted no effect on luciferase activity of BCL2L11-Mut in primary hippocampal neurons. (C) Silencing miR-338-5p caused a significant increase in luciferase activity of BCL2L11-WT and exerted no effect on luciferase activity of BCL2L11-Mut in primary hippocampal neurons. (D–F) Relative BCL2L11 mRNA level (D) and protein expression (E, F) of primary hippocampal neurons transfected with NC mimic or miR-338-5p mimic, or NC antagomir or miR-338-5p antagomir determined by qRT-PCR and Western blot respectively. (G–I) Primary hippocampal neurons were transfected with pcDNA3.1-vector or pcDNA3.1-BCL2L11. Two days later, neurons were cultured for consecutive 7 days with or without 5 mM Aβ40. (G, H) The representative immunofluorescent images (G) and quantification (H) of TUNEL-positive hippocampal neurons in vitro. (I, J) The quantification of TUNEL-positive hippocampal neuron in vitro. The representative western blot images (I) and quantification analysis (J) of cleaved caspase-3 expression. Scale bar=50 μm. Results are presented as mean ± SD. n = 3 in each group. *P < 0.05; **P<0.01.