![Fig. 6. Thrombin stimulation of Egr-1 mRNA in primary endothelial cells is mediated by an ERK1/2 MAPK–dependent pathway. / HPAECs were starved overnight in EBM-2 medium containing 0.5% FBS and then treated with or without 1.5 U/mL thrombin for 1 hour. Alternatively, the serum-starved cells were preincubated with 20 μM PD98059 (PD), 3 μM SB203580 (SB), 5 μM BIM, 10 μM LY294002 (LY), or dimethyl sulfoxide (DMSO) (1 μL as solvent control) for 10 minutes and then incubated for 1 hour with medium containing no thrombin or 1.5 U/mL thrombin. RNase protection assays were carried out with α-[32P] UTP-labeled Egr-1 and β-actin probes as described in “Materials and methods.” Densitometry was used to calculate the ratio of Egr-1 and β-actin signals. The results are expressed as fold induction relative to control untreated cells. The data represent the means and standard deviations from 3 independent experiments.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/100/13/10.1182_blood-2002-02-0415/4/h82423529006.jpeg?Expires=1735507425&Signature=j-J8ulBqUBidn9g3IPZZN8sf0~dnTwLGvqYDP44JUA62oH9JR82heXs7g7misTF22gi7ors9zaf7kek7jdn3r9ow4uCn70UKNEIHG7DMlTOCtCIzgZQdcbnBPwu7tQl4aSbnq8COFzY5u3FqQtgAGXdoOaRASkzUO~Q6de~CefQHo4frtsuC3eB-b8I4vAsQDB1Axwb9QrWtV15rw7T8SFJc4sFASLfLMLf2zW~6~XlyD9QdnYvyqeMruGGPPJXMEDaE9MF3XteCAOVVpWWTviizaKEkVmLdXCclpcip4SnHFezuI1B3lpuJPd-IKfBU0mDcj58MzcpwnVadXBT4kg__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Thrombin stimulation of Egr-1 mRNA in primary endothelial cells is mediated by an ERK1/2 MAPK–dependent pathway.
HPAECs were starved overnight in EBM-2 medium containing 0.5% FBS and then treated with or without 1.5 U/mL thrombin for 1 hour. Alternatively, the serum-starved cells were preincubated with 20 μM PD98059 (PD), 3 μM SB203580 (SB), 5 μM BIM, 10 μM LY294002 (LY), or dimethyl sulfoxide (DMSO) (1 μL as solvent control) for 10 minutes and then incubated for 1 hour with medium containing no thrombin or 1.5 U/mL thrombin. RNase protection assays were carried out with α-[32P] UTP-labeled Egr-1 and β-actin probes as described in “Materials and methods.” Densitometry was used to calculate the ratio of Egr-1 and β-actin signals. The results are expressed as fold induction relative to control untreated cells. The data represent the means and standard deviations from 3 independent experiments.
