Gq-coupled muscarinic receptor-induced activation of the ERK5 pathway does not require receptor phosphorylation (original) (raw)

2013

Abstract

Different stable lines of CHO cells (previously characterised in [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084174#B11" target="_blank">11</a>]) that stably overexpress wild-type or phosphorylation-deficient muscarinic M3 receptor (A) and wild-type or phosphorylation-deficient muscarinic M1 receptor (B) were utilised. All cell lines were transfected with HA-ERK5. Twenty-four hours after transfection, cells were serum-starved for 2h and stimulated with acetylcholine (100µM) for the indicated times (A) or incubated for 15min with acetylcholine at various concentrations (B). ERK5 phosphorylation was assessed as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084174#pone-0084174-g002" target="_blank">Figure 2</a>. Data (mean +/- SD of 2-3 independent experiments) were normalised using HA-ERK5 as loading control and expressed as fold-induction over basal conditions or over maximum activation (*p<0.05, **p<0.005; two-tailed T-test). </p

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