Figure 9. Examination of the binding of ZFP36L1-containing protein complexes on Pparγ2 mRNA.

A. Western blot analyses. The ZFP36L1 levels of C3H10T1/2 cells overexpressing Flag-tagged ZFP36L1 (F-ZFP36L1) and control cells (F-EV) were shown. B. RT-qPCR analyses. F-EV and F-ZFP36L1 cells were induced to undergo adipogenic differentiation with or without concomitant treatment of troglitazone (TZD, 1 μM) for 24h. The expression of Pparγ2 mRNA was examined. C. Ribonucleoprotein immunoprecipitation and RT-qPCR analyses. One milligram of lysates prepared from F-EV or F-ZFP36L1 cells (1 × 10⁷) were incubated with protein A beads precoated with 15 μg of either anti-Flag or anti-IgG antibody to precipitate ribonucleoprotein complexes and to extract RNAs from the complexes as described in Materials and Methods. RNAs were used in subsequent RT-qPCR assays for Pparγ2 and β-actin mRNAs. Pparγ2 signals were normalized to β-actin signals. The normalized Pparγ2 signals obtained from the ribonucleoprotein complexes pulled down by anti-Flag antibody were compared with those pulled down by anti-IgG antibody (to which a value of 1 was assigned). Data represent the mean ± S.D. from three analyses. *, P < 0.05.