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Fig. 6. Tyrosine dephosphorylation of endogenous Kir2.1 channels at the onset of
differentiation. (A) Myoblasts (300x106) were
cultured in growth medium (GM) or in differentiation medium for 6 hours (DM
6h). Both populations of myoblasts were treated for 30 minutes preceding lysis
with 10 µM bpV(Phen) to avoid unspecific dephosphorylation. An equal amount
of proteins (10 mg) from each population was immunoprecipitated with a rabbit
anti-Kir2.1 antibody. Immunoprecipitated proteins were separated on SDS-PAGE,
and revealed with chicken anti-Kir2.1 antibody (upper lane) and then reblotted
with an anti-phosphotyrosine antibody (PT-66, lower lane). Bands were
quantified by Optiquant software. (B) The histogram represents the
fraction of endogenous Kir2.1 channels that are tyrosine-phosphorylated
normalized to the tyrosine-phosphorylated Kir2.1 channels in proliferating
conditions. Results were obtained from three independent experiments.
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