Figure 7.

Functional analysis of ZFP36L1 and ZFP36L2 in adipogenesis. (A) Expression profiles of TTP family proteins and mRNAs during adipogenesis. Two-day post-confluent 3T3-L1 cells were induced to differentiate for 1 to 5 days, and WCEs were prepared at the indicated times and immunoblotted with anti-TTP, anti-ZFP36L1, anti-ZFP36L2. Tubulin served as a loading control (left panel). RNA was isolated at indicated time for quantitative PCR (right panel). (B) The effects of ZFP36L1 and ZFP36L2 knockdown on adipogenesis. Lentivirus was used to infect 3T3-L1 with shRNA against mouse ZFP36L1, ZFP36L2, or luciferase (control). Infected 3T3-L1 cells were induced to differentiate for 6 days. The cells were stained with oil red O and observed with microscope, scale bar, 50 μm (right panel). Oil red O was extracted using isopropanol, and OD₅₁₀ was measured to quantify triglyceride accumulation (left panel). The lower panel shows the expression of the adipogenic marker aP2 after differentiation for 0 to 6 days. *P < 0.05, **P < 0.01. (C) The effect of TTP and MKP-1 knockdown on adipogenesis. 3T3-L1 cells were transfected with siRNA against TTP, MKP-1, or non-targeting control. After induction for 6 days, the cells were stained with oil red O to quantify the differentiation. The graph shows the mean ± SD of relative OD₅₁₀ from three independent experiments. **P < 0.01.