Fig. 6

miR-146a-5p reduces proteasomal degradation of DVL2 via silencing the NEDD4L expression and modulates Wnt/β-catenin signaling (n = 3). (A-C) The protein expression levels of NEDD4L and DVL2 in IPEC-J2 cells were transfected with pcDNA3.1-NEDD4L or pcDNA3.1-control, then treated with MG132 or DMSO, and followed by Western blot analysis. (D-G) The protein expression levels of NEDD4L, DVL2, and β-catenin in IPEC-J2 cells were transfected with miR-146a-5p mimics or NC, then treated with MG132 or DMSO, and followed by Western blot analysis. (H-J) The relative TOP/FOP Flash reporter activities of IPEC-J2 cells after transfected with miR-146a-5p mimics or NC (H), pcDNA3.1-NEDD4L or pcDNA3.1-control (I), pcDNA3.1-DVL2 or pcDNA3.1-control (J). (K) Western blot images of HEK293 cells were transfected with constructs expressing Flag-NEDD4L, HA-Ubiquitin (HA-UB), MYC-DVL2, and miR-146-5p mimics or mimics-NC, then followed by anti-MYC IP under denaturing conditions. Data are expressed as means ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001