Research Paper Volume 10, Issue 12 pp 3794—3805

Sevoflurane impairs learning and memory of the developing brain through post-transcriptional inhibition of CCNA2 via microRNA-19-3p

Figure 4. MiR-19-3p targets CCNA2 to suppress its translation in neural cells. (A) MiR-19-3p was overexpressed by a plasmid carrying miR-19-3p or knocked down by a plasmid carrying as-miR-19-3p in a human neural cell line, HCN-2. Transfection with a null sequence was used as a control. RT-qPCR for miR-19-3p levels in these cells. (B) An intact 3'-UTR of CCNA2 mRNA (CCNA2 3'-UTR) and an 3'-UTR of CCNA2 mRNA with a mutant at miR-19-3p-binding site (CCNA2 3'-UTR mut) were prepared. A dual luciferase reporter assay was performed using combinations of one miR-19-3p-modifying plasmid and one CCNA2 3’-UTR plasmid. (C-D) RT-qPCR (C) and Western blot (D) for CCNA2 levels in miR-19-3p-modified HCN-2 cells. *p<0.05. NS: non-significant. N=5.