Figure 5.

MIR4458, MIR4667-5p, and MIR4668-5p target and suppress ATG10. (A) Sequences of MIR4458, MIR4667-5p, and MIR4668-5p, and the potential binding site at the 3′UTR of ATG10. Also shown are nucleotides mutated in the ATG10-3′UTR mutant. (B) Luciferase reporter assay. HEK293 cells were transiently cotransfected miRNA mimics or control with luciferase reporter vectors. Luciferase activity was normalized to the activity of Renilla luciferase. (C and D) Western blot analysis of endogenous ATG10 in uninfected and B. pseudomallei-infected A549 cells after transfected with miRNA control, mimics, or inhibitors. (E and G) The mRNA level of ATG10 and the protein levels of ATG10 were determined by qRT-PCR and western blot. HEK293 cells were cotransfected the ATG10-expressing plasmid containing wild-type 3′UTR or the control plasmid, with miRNA mimics or control for 24 h. Data are representative of at least 3 independent experiments (*P < 0.05, **P < 0.01).