Figure 8 | Scientific Reports

Figure 8

From: Receptor tyrosine kinase EphA7 is required for interneuron connectivity at specific subcellular compartments of granule cells

Figure 8

Differential interactions of activated mTOR with gephyrin and collybistin.

(a,b) Cortical neurons were treated with ephrin A5 or BDNF for mTOR stimulation. Cell lysates were prepared for subsequent immunoprecipitation of endogenous mTOR and analysis of coprecipitated gephyrin after western blotting. Decreased mTOR-gephyrin interactions were observed after mTOR stimulation. (b) Densitometric quantification of bands depicted in (a) and determination of the gephyrin/mTOR ratio; n = 3, ANOVA, Fisher-PLSD; error bars, S.E.M. (c–f) Expression of ca-mTOR decreased EGFP-gephyrin/mTOR and increased collybistin/mTOR interactions. HeLa cells were transfected with expression vectors for wild type (WT) or constitutively active (CA) mTOR, EGFP-gephyrin (Geph) and myc-tagged collybistin (myc-CB). In some samples plasmid pUC18 (pUC) was added for equal amounts of transfected DNA. EGFP-gephyrin levels were related to the corresponding mTOR or collybistin signals. mTOR-gephyrin coprecipitation; n = 3; ***p < 0.0001. mTOR-myc-collybistin; n = 4; **p < 0.01; Kruskal-Wallis-Test, followed by Tukey-Test; error bars, S.E.M.

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