![Figure 1. Induction of PAI-1 gene expression by A23187 and BAPTA-am. (A) Measurement of intracellular cellular calcium [Ca2+]i. Cultured HepG2 cells were treated with Fura-2 am (10 μM) for 1 hour under normoxia (16% O2) and further cultured under normoxia or hypoxia (8% O2) with A23187 (5 μM), BAPTA-am (5 μM), or BAPTA (5 μM) for 2 hours. In each experiment [Ca2+]i at 16% O2 was set to 100%. Values are means ± SEM of 3 independent culture experiments. Statistics, Student t test for paired values: P ≤ .05, compared with 16% O2. (B) HepG2 cells were treated with A23187 or BAPTA-am as in panel A and further cultured under normoxia (16% O2) or hypoxia (8% O2). The PAI-1 mRNA measured after 4 hours or the PAI-1 protein level measured after 24 hours under hypoxia was set to 100%. Values are means ± SEM of 3 independent culture experiments. Statistics, Student t test for paired values: *P ≤ .05, compared with the control at the same oxygen tension (pO2). (C) Representative Northern and Western blots. For Northern analysis 15 μg total RNA was hybridized to digoxigenin-labeled PAI-1 and β-actin antisense RNA probes (described in “Materials and methods”). Protein (50 μg) from the medium was subjected to Western analysis with an antibody against human PAI-1 or albumin, respectively. Autoradiographic signals were obtained by chemiluminescence and scanned by videodensitometry.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/104/13/10.1182_blood-2004-03-1017/6/zh80240470700001.jpeg?Expires=1724263095&Signature=BJjHLW~ltG1Zwoufg-9HliTuTYvf7iGcuTQeBhw7jFGqJxMbfDH3gxT9bZxPYlinVy6F2geAX2SoHwgzldKGXep54VYE7YdMud7Hm8PT07eeHwaNAD7a8WHcZNgHrOHpHtAl4VwBJNoTDLOGNa9jcX9Glzugf6SOnlzuUGotI8qVLB0WRzXsdccLrOrG1ggQz9EoEc-0Nuo4NLQtvYUcoLB9Med3bd7L4hZv4A2QTlQWEWR-u~AnMumCLu8skgDaDow8SRddEuus-5bLX3lSvAdSS92h3StxH9TkkxKs0LIh4VHmP4S7VehoRPnO6d7y35PeU32W7HS0m~x--z0LIQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Induction of PAI-1 gene expression by A23187 and BAPTA-am. (A) Measurement of intracellular cellular calcium [Ca2+]i. Cultured HepG2 cells were treated with Fura-2 am (10 μM) for 1 hour under normoxia (16% O2) and further cultured under normoxia or hypoxia (8% O2) with A23187 (5 μM), BAPTA-am (5 μM), or BAPTA (5 μM) for 2 hours. In each experiment [Ca2+]i at 16% O2 was set to 100%. Values are means ± SEM of 3 independent culture experiments. Statistics, Student t test for paired values: P ≤ .05, compared with 16% O2. (B) HepG2 cells were treated with A23187 or BAPTA-am as in panel A and further cultured under normoxia (16% O2) or hypoxia (8% O2). The PAI-1 mRNA measured after 4 hours or the PAI-1 protein level measured after 24 hours under hypoxia was set to 100%. Values are means ± SEM of 3 independent culture experiments. Statistics, Student t test for paired values: *P ≤ .05, compared with the control at the same oxygen tension (pO2). (C) Representative Northern and Western blots. For Northern analysis 15 μg total RNA was hybridized to digoxigenin-labeled PAI-1 and β-actin antisense RNA probes (described in “Materials and methods”). Protein (50 μg) from the medium was subjected to Western analysis with an antibody against human PAI-1 or albumin, respectively. Autoradiographic signals were obtained by chemiluminescence and scanned by videodensitometry.
