Journal: Journal of immunology (Baltimore, Md. : 1950)
Article Title: Alpha1-Antitrypsin combines with plasma fatty acids and induces angiopoietin-like protein 4 expression
Figure Lengend Snippet: A1AT-LA induced expression of Angptl4 and FABP4 is related to PPARγ and PPARβ/δ activity. Cells were pretreated for 30 min with 10 μM GW9662, an irreversible PPARγ antagonist (A, B), or with 1 μM ST247, a selective and inverse agonist of PPARβ/δ (C, D), prior to addition of 1 mg/ml A1AT-0, A1AT-LA, LA preparation or Prolastin® for another 6 h. Expression levels of Angptl4 (A, C) and FABP4 (B, D) were determined by RT-PCR and normalized to GAPDH. In (A, B) each point represents mean ±SD of n=3 independent experiments, each with 3 repeats. In (C, D) each point represents mean ±SD of n=2 independent experiments, each with 3 repeats.
Article Snippet: For specific detection, the following primary antibodies were used: rabbit polyclonal anti A1AT (DAKO), mouse monoclonal anti A1AT (clone B9, Santa Cruz Biotechnology), mouse monoclonal anti polymeric A1AT (Clone 2C1, Hycult biotech), mouse monoclonal anti FABP4 (clone 1105CT1-1-1, Antibodies-Online GmbH), rabbit polyclonal anti ERK1/2 and mouse monoclonal anti p-ERK1/2 (both from Sigma-Aldrich), monoclonal anti β-actin (AC-15, Sigma-Aldrich).
Techniques: Expressing, Activity Assay, Reverse Transcription Polymerase Chain Reaction