Fig. 3
miR-30c-1-3p directly targets ATG4B and ATG9B. (A) Targetscan predicts miR-30c-1-3p interaction sequence with ATG4B and ATG9B. (B, C) miR-30c-1-3p mimics regulation of ATG4B and ATG9B 3’-UTRs reporter. The luciferase reporter assay co-transfected the pmirGLO dual luciferase reporter vector (WT, Mut, and Blank) and miR-30c-1-3p mimics or mimics nc 24 h. (D) The expression of miR-30c-1-3p was observed in THP-1 transfected with miR-30c-1-3p mimics and mimics nc 24 h. (E) Western Blot detection of ATG4B and ATG9B expression in THP-1 transfected with miR-30c-1-3p mimics and mimics nc 24 h. The ATG4B and ATG9B bands were quantified relative to β-actin. (F) Fluorescent microscopic imaging revealed that the expression of GFP in the pEGFP-ATG4B and pEGFP-ATG9B reporters was inhibited by miR-30c-1-3p. THP-1 cells were transfected with GFP reporter vectors (WT and Mut) and fluorescence intensity was subsequently detected by immunofluorescence staining following transfection with miR-30c-1-3p mimics and mimics nc for 24 h. Scale bars: 10 μm. All data above represent the means ± SD of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not statistically different.
