Figure 6
miR-124 targets TACE to inhibit TNF-α production. (A) Western blotting of pro-TNF-α and TNF-α proteins in RAW264.7 cells incubated with 5 mM nicotine for 10 min or transfected with miR-124 mimic for 36 h, and then stimulated with 100 ng/ml LPS for 9 h. (B) Luciferase activity of HEK293 cells co-transfected with the miR-124 mimic or inhibitor and the luciferase reporter of TACE 3′-UTR wild type (WT) or Mutant (Mut) for 24 h. (C) Western blotting of TACE protein in RAW264.7 cells treated as described in A. The two bands detected may represent processing or glycosylation of TACE as indicated by the Ab manufacturer. (D) ELISA detection of TNF-α in the supernatants of RAW264.7 cells transfected with TACE siRNA and miR-124 mimic for 36 h and then treated with 100 ng/ml LPS for 9 h. For A and C, data are representative of three independent experiments. For B and D, data represent mean ± SD (n = 3). NS, not significant; **P < 0.01 by Student's t-test.
