Fig. 4

UBE2K is a target of miR-3968. a Venn diagram illustrates the intersection of predicted target genes of miR-3968 from three distinct online tools. b The biological processes (BP) and molecular function (MF) GO terms for the 21 intersected genes (depicted in a) were predicted by clusterProfiler R package (Version 4.8.3). c Two different predicted sites of miR-3968 binding within the 3′-UTR of UBE2K and two distinct mutant sequences of the UBE2K 3′-UTR were constructed. Dual-luciferase reporter assays revealed that miR-3968 mimic could only suppress the luciferase activity of WT 3′-UTR of UBE2K in HEK293 cells (n = 3). d The mRNA levels of miR-3968 and UBE2K were detected by qRT-PCR in N2a cells transfected with miR-3968 mimic (Mimic), inhibitor (Inhibitor), or the corresponding scramble control (Scramble) (n = 4). e The mRNA level of UBE2K in N2a cells transfected with circRIMS2 (n = 4). f The protein levels of UBE2K in N2a cells with transfection of Mimic or Inhibitor were detected by WB (n = 3). g qRT-PCR analysis of UBE2K in the hippocampus of APP/PS1 and WT mice at diverse stages (n = 4). h The protein levels of UBE2K in the hippocampus of WT and APP/PS1 mice at different stages were detected by WB (n = 4). Data are presented as mean ± S.E.M. and two-tailed t tests were used unless otherwise specified. *P < 0.05, **P < 0.01, ***P < 0.001