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Figure 1 | Molecular Brain

Figure 1

From: Pyk2 uncouples metabotropic glutamate receptor G protein signaling but facilitates ERK1/2 activation

Figure 1

Pyk2 interacts with Group I mGluRs. A) Representative immunoblot showing the co-immunoprecipitation of HA-Pyk2 with Flag-mGluR1a from HEK293 treated with 30 μM quisqualate for 0, 10, and 20 min. FLAG-mGluR1a is immunopreciptated from 500-1000 μg of protein lysate. The expression of HA-Pyk2 is shown in input lanes correspond to 100 μg of protein lysate. B) The densitometric analysis of autoradiographs showing the mean ± SD of 5 independent experiments examining the co-immunoprecipitation of HA-Pyk2 with FLAG-mGluR5. * P < 0.05 versus untreated cells. C) Representative immunoblotblot showing the co-immunoprecipitation of Pyk2 with mGluR1 and mGluR5 polyclonal antibodies from either 1000 or 500 μg of rat whole brain lysates. The blot is representative of 3 independent experiments. D) Representative immunoblot of HA-Pyk2 co-precipitated with 1 μg of GST, and GST fusion proteins encoding either the second intracellular loop domain of mGluR1 (GST-IL2) or the distal region of the carboxyl-terminal tail domain of mGluR1a (GST-CTC). The GST fusions are incubated with 400 μg of COS-7 cell lysates expressing HA-Pyk2. The expression of HA-Pyk2 is shown in input lanes correspond to 100 μg of protein lysate. E) Representative immunoblot showing the co-immunoprecipitation of HA-Pyk2 with either Flag-mGluR1a-866Δ or FLAG-mGluR1b and HA-Pyk2 and treated with 30 μM quisqualate for 0 and 30 min. FLAG-mGluR1 protein is immunopreciptated from 500-1000 μg of protein lysate. The expression of HA-Pyk2 is shown in input lanes correspond to 100 μg of protein lysate. Data are representative of 4 independent experiments.

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