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Example sentences for: wortmannin
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Two inhibitors of PI 3-kinase, wortmannin and LY294002, and an inhibitor of p70 S6 kinase activation, rapamycin, blocked the inhibitory effect of insulin on SP-A gene expression.
Wortmannin is a fungal metabolite that is a relatively selective inhibitor of PI 3-kinase with a half-maximal inhibition (IC 50 ) less than 5 nM [ 11 ] . In the wortmannin experiments, media were changed every 2 hours for a total of 12 hours in all of the conditions because wortmannin is unstable in solution at physiological pH [ 19 ] . The shorter incubation time with insulin in the wortmannin experiments resulted in a decreased inhibitory effect of insulin, i.e., an ~22% inhibition of SP-A mRNA levels (Fig.
H441 cells resemble bronchiolar epithelial Clara cells in phenotype and produce both SP-A and SP-B mRNA and protein [ 14 ] . The hormonal regulation of SP-A and SP-B gene expression in H441 cells is similar to that observed in differentiated type II cells in human fetal lung explants [ 6 7 15 16 ] . We treated H441 cells with the four inhibitors of signal transduction: wortmannin, LY294002, rapamycin and PD98059, to elucidate which signaling pathways are activated when insulin inhibits SP-A gene expression.
Wortmannin, LY294002, rapamycin and PD98059 were reconstituted in dimethyl sulphoxide as 1 mM, 50 mM, 50 μM and 10 mM stock solutions, respectively, and stored at -80°C in aliquots.
Wortmannin, added alone, did not significantly alter SP-A mRNA levels at any concentration tested (Fig.