Fig. 5

miR-145a-5p suppressed inflammation by targeting TLR4/ NF-κB signaling in vitro. (A) Schematic diagrams showing the predicted binding sequence between miR-145a-5p and wide-type (WT) or mutant 3’-UTRs of TLR4. (B) Dual luciferase assay was performed in HEK293T cells after co-transfected with reporter plasmids containing WT or mutant 3’-UTRs of TLR4 as well as miR-145a-5p mimics or Ctrl (n = 4). (C) Representative Western blot showing the expression of TLR4 and NF-kB signal-related molecules such as p65 in BV2 that were transfected with miR-145a-5p mimics or Ctrl followed by PBS, LPS + IFN-γ or IL-4 stimulation for 24 h. (D) Quantitative analysis of the protein level of TLR4 and p65 as well as P-p65 in (D) (n = 4). (E) BV2 was transfected with miR-145a-5p inhibitor or Ctrl and siTLR4 followed by LPS + IFN-γ stimulation for 24 h. The expression of TLR4, p65 and P-p65, as well as TNF-α, IL-1β and iNOS was detected by Western blot (n = 4). (F) Quantitative analysis of the protein level in (E) (n = 4). Data shown as mean ± SEM, *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001 by one-way ANOVA with Turkey’s multiple comparison test