Thrombin activates a Y box–binding protein (DNA-binding protein B) in endothelial cells
J. Clin. Invest. Olga I. Stenina, et al. 106:579 doi:10.1172/JCI9075 [Go to this article.]

Figure 5
Effect of tyrosine phosphatase inhibitors on activation of dbpB and PDGF B–chain transcription. (a) Effect of sodium vanadate (Na₃VO₄) and sodium fluoride (NaF) on dbpB activation by thrombin. ECs were pretreated with Na₃VO₄ or NaF (400 μM for 30 minutes) and then stimulated with thrombin (10 U/mL for 2 hours). EMSA was performed as described in Methods. Control, extract from untreated EC; Thr, extract from EC stimulated with thrombin. (b) Effect of sodium vanadate (Na₃VO₄) and phenyl arsine oxide (PAO) on thrombin-induced PDGF B–chain mRNA in ECs. ECs were pretreated with Na₃VO₄ or PAO and then stimulated with thrombin (10 U/mL for 6 hours). Total RNA was extracted using Trizol reagent and Northern hybridization was performed using either the PDGF B–chain or GAPDH cDNAs as probe. (c) Effect of PTP inhibitors on thrombin-induced transcription driven by the PDGF B–chain promoter. Bovine ECs were transiently transfected (as described in Methods) with a luciferase reporter gene under the control of a 400-bp PDGF B–chain promoter fragment in pGL3-Basic vector (Promega Corp.). Pretreatment (30 minutes) with PTP inhibitors (400 μM Na₃VO_4, or 200 nM PAO) was initiated after a 6-hour transfection incubation. Lysates were prepared after a 15-hour incubation with thrombin (10 U/mL). Luciferase activity was normalized to β-galactosidase activity derived from cotransfected pSV β-galactosidase cDNA.