Figure 6.

miR-16 was a direct target of TLR4 and regulated NF-κB signaling

(a) There are predicted binding sites between miR-16 and TLR4. (b) BV-2 cells were transfected with miR-16 mimic or mimic-NC, and the expression level of miR-16 was detected using quantitative real-time PCR. (c) The binding relationship between miR-16 and TLR4 was verified by luciferase reporter assay. *** p < 0.001 vs mimic-NC. (d) BV-2 cells were pretreated with DHA with or without transfection with inhibitor-NC or miR-16 inhibitor, prior to morphine induction. The protein expression of TLR4 was detected by western blot, and the relative expression level was quantified. (e) The expression of NF-κB p65 was measured using immunofluorescence assay. *** p < 0.001 vs control group; ^### p < 0.001 vs morphine group; ^ΔΔΔ p < 0.001 vs morphine + DHA + inhibitor NC group.