Figure 4.

miR-362-3p targets MyD88. The binding sites and mutation location (positions 20–26) are listed (A). miR-362-3p mimics and a plasmid containing either a wild-type or mutated 3′-UTR sequence of MyD88 were transfected into SKOV3 cells. After 48 h, the luciferase activity was analyzed (B). miR-362-3p mimics were transfected into SKOV3 cells and the amount of MyD88 protein was determined via western blot (C). MyD88 levels in 12 OC tissues and their matched adjacent normal tissues were assessed via qRT-PCR. The relative MyD88 levels tumor/normal (log2) are listed (D). Pearson's correlation coefficient analysis revealed that miR-362-3p levels and MyD88 mRNA levels were inversely correlated in the OC tissue samples (E). SKOV3 and HO8910 cells (6×10⁵ cells/well) were transfected with miR-362-3p or miR-NC (NC) separately, followed by transfection 12 h later with an empty plasmid or a MyD88-overexpression plasmid (pcDNA3.1-MyD88). The western blot was performed to test the MyD88 protein levels following transfection (F). An MTT assay was performed, and the OD value is presented as the relative cell number. The relative cell number in the miR-NC + vector (negative control) was defined as l00% (G). The full, not adjusted blot image for figure 4C and figure-4F were include as supplementary materials. (SM1: figure-4C β-actin, SM2: figure-4C MyD88, SM3: figure-4F β-actin, SM4: figure-4F MyD88). Each experiment was repeated at least three times. ∗P < 0.05.