Figure 3. Regulation of Dicer Expression in 3T3-F442A Preadipocytes in Response to Stress.

(A) 3T3-F442A preadipocytes were treated with sub-lethal doses of different stress agents (1 mM H₂O₂, 50 mU/mL glucose oxidase). Dicer mRNA expression was assessed by RT-qPCR at the indicated time-points. After 90 min, medium containing H₂O₂ was replaced by H₂O₂-free medium. Glucose oxidase was maintained in the medium throughout the entire experiment. Experiments were repeated twice in duplicate.

(B) Dicer expression was analyzed by Western blotting in 3T3-F442A preadipocytes in response to 1 mM H₂O₂ for 3h.

(C) Dose-response of Dicer mRNA expression to different stress agents in 3T3-F442A preadipocytes as measured by RT-qPCR. Time-points were 1.5h for H₂O₂, 3h for glucose oxidase and UV, and 24h for paraquat. (n=4 per group).

(D) 3T3-F442A preadipocytes were treated with sub-lethal doses of different stress agents (2 μg/mL tunicamycin, 25 ng/mL TNFα, 100 ng/mL Fas ligand) and Dicer mRNA expression was assessed by RT-qPCR at the indicated time-points. Experiments were repeated twice in duplicate.

(E) 3T3-F442A preadipocytes were pre-treated with 150 nM Insulin for 1h, followed by treatment with 1 mM H₂O₂ for 90 min. Dicer mRNA expression was determined by RT-qPCR. (n=4 per group).

(F) 3T3-F442A preadipocytes were serum starved (6h) or treated with agents to mimic nutrient deprivation (1 mM AICAR for 6h and 1 μM rapamycin for 3h), and Dicer mRNA expression was assessed by RT-qPCR. Experiments were repeated twice in duplicate.

* P < 0.05. Error bars, SEM.