Figure 5

IL-17 enhances gene expression by modulating AUF1-mediated mRNA decay in MSCs. (a–c) Wild-type (a) or auf1^−/− MSCs (b), shCTRL MSCs or shAUF1 MSCs (c) were treated with IFNγ and TNFα, with or without IL-17 (all cytokines supplemented at 10 ng/ml) for 6 h, and then actinomycin D (5 μg/ml) was added to stop transcription. At the indicated time points, mRNA levels were assayed by quantitative RT-PCR, taking the expression level at the time of actinomycin D addition as 100%. mRNA expression values are means±S.E.M. of three wells from a representative of three independent experiments. (d) Wild-type MSCs were treated with IFNγ and TNFα, with or without IL-17 (all cytokines supplemented at 10 ng/ml). At different time points, cells were collected for analysis of AUF1 protein level by western blot. (e) Wild-type MSCs were treated with IL-17 (10 ng/ml) for different time, and cells were harvested for analysis of interaction of Act1 and AUF1 by immunoprecipitation. Western blotting data is from a representative of two independent experiments