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 2
Pin1 associates with and is downstream of PKC-α. (A and C) RT-qPCR analysis for TGF-β1 mRNA. (A) Eos were treated as in Figure 1A. Gö, Gö6976; SA, safingol; PD, PD98059 (50 μM). (B) ELISA for TGF-β1 protein. Cells were treated as in Figure 1D. (C) Eos were treated as in A. HETE, 12(S)-HETE (10 nM/ml); TXA, thymeleatoxin (10 nM). (D) Eos were treated for 4 hours with HA alone or with juglone. Lysates were immunoprecipitated with anti-Pin1 followed by immunoblotting (as shown on right). Input, 10% of lysates before IP. The blot represents 1 of 2 independent experiments. (E and F) Pin1 isomerase assay of cytoplasmic lysates from untreated Eos (R) or Eos treated for 10 minutes with HA alone, with juglone, or with Gö6976 (HA+Gö) (E) or with 12(S)-HETE alone or with Gö6976 (HETE+Gö) (F). The kinetics (left) is representative of 3 independent experiments. The released Δ[pNA]/g of protein was calculated (right), as described (20). (G) Immunoblot of cell lysates from Eos treated for 4 hours with HA alone or with juglone or with 50 μM of MG132 (HA+J+MG). The blot represents 1 of 2 independent experiments. 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.