Words similar to cos-
Example sentences for: cos-
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COS-7 cells were transfected in 6 cm plates with 3 μg of the indicated expression plasmid.
Initially, we expressed the Src KM/Cas(SD) (entire Cas SD) in Cos-7 cells alone, or along with wild type (WT) v-crk, or v-crk SH2 or SH3 mutants and assayed for tyrosine phosphorylation and association by co-immunoprecipitation (Fig.
Two molecular loci recently have been implicated as potential modulators of GnRH receptor signaling: one is the receptor itself [ 23 ] , and the other is the G protein that transduces the signal generated by binding of the receptor to its agonist [ 9 ] . With respect to the former locus, G protein coupled receptor kinases (GRKs) act in an agonist-specific manner to phosphorylate intracellular regions of the receptor thus permitting β arrestin binding that sterically inhibits G protein association with the receptor [ 1 45 ] . We have reported previously that experimental expression of GRKs in GnRH receptor-expressing heterologous cells (COS-1) suppressed GnRH-stimulated IP 3 production, and that co-expression of GRK2 and β-arrestin 2 suppressed GnRH receptor signaling more than that of either alone [ 23 ] . Stronger evidence that the GRK/β arrestin paradigm may play a role in regulating GnRH receptor signaling is our recent finding that adenovirus-mediated gene transfer of GRK2 into normal pituitary gonadotropes suppressed GnRH-stimulated LH secretion and IP 3 production [ 31 ] . However, no evidence has been presented for a direct interaction between β-arrestin and the GnRH receptor.
With respect to the G protein as a locus for modulation of GnRH receptor signaling, we reported earlier that experimental expression of RGS3 but not RGS1, 2, or 4 in GnRH receptor expressing COS-1 cells suppressed GnRH-stimulated IP 3 production [ 9 ] . In the current report, we present evidence strengthening the hypothesis of a regulatory role for RGS3 in GnRH receptor signaling: adenovirus-mediated expression of RGS3 in normal pituitary gonadotropes profoundly inhibited GnRH-stimulated LH secretion and 3H-inositol phosphate accumulation.
YZ4 was then subcloned into the eukaryotic expression vector pcDNA3 (Invitrogen) and the resultant plasmid, pcDNA3/REP 2 , was transfected into COS-1 using lipofectamine according to the manufacturer's directions (Life Science Technologies) with 12 μg plasmid DNA and 45 μL lipofectamine solution.
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