Figure 7.

MiR-124 failed to promote degradation TNF-α protein when USP2 and USP14 were both knocked down. RAW264.7 cells were transfected control RNAi or USP2 and USP14 RNAi in the absence or presence of miR-124 for 36 h. For co-transfection, RAW264.7 cells were first transfected with USP2 and USP14 RNAi for 16 h, then transfected with miR-124 mimic for another 24 h. Cells were then stimulated with 100 ng/mL of LPS for 9 h, followed by incubation with 30 μg/mL of CHX for the time indicated. TNF-α protein in the cells (A) and in the supernatant (B) was detected and analyzed as described in Figure 3. Mean±SD. (n=3). ^**P<0.01 control. ^##P<0.01 USP2/14 RNAi+miR-124 vs USP2/USP14 RNAi.