Pin1 regulates TGF-β1 production by activated human and murine eosinophils and contributes to allergic lung fibrosis
J. Clin. Invest. Zhong-Jian Shen, et al. 118:479 doi:10.1172/JCI32789 [Go to this article.]

Figure 1
Pin1 is required for TGF-β1 mRNA expression. (A, B, C, and E) Reverse transcription and qPCR (RT-qPCR) analysis for TGF-β1 mRNA. (A) Eos left untreated (resting [R]) or incubated for 4 hours with HA alone (100 μg/ml in all experiments) or with juglone (0.1 or 1.0 μM) prior to analysis. (B) Cells incubated for 4 hours with HA alone, with 60–300 nM TAT-WW-Pin1 (WW) or TAT-GFP (GFP) prior to analysis. (C) Cells left untreated or incubated for the indicated times with HA alone or with juglone (1.0 μM) (HA+J) or juglone alone (J) before collection and analysis at the times shown. (D) Intracellular levels of TGF-β1 protein were measured by immunoblotting of total cell lysates under reducing conditions and signal quantitated by densitometry and normalized to β-actin. (E) ELISA for TGF-β1 protein. Cells left untreated or incubated for 24 hours with HA alone or with juglone before collection of culture medium. The data are mean of 2 independent experiments. (F) Decay of TGF-β1 mRNA. Cells were treated for 2 hours with HA alone or with juglone, followed by the addition of 50 μg/ml of DRB (5,6-dichloro-1-β-d-ribofuranosylbenzimidazole) to block transcription. Cells were then collected at the times shown for analysis. Error bars indicate mean ± SD of 3 independent experiments with different donors. *P < 0.05 by Student’s t test in a 2-tailed analysis.