Figure 3.

Identification of Interaction between GABA_AR and Hyperekplexic Mutant GlyRs

(A) GlyRα₁ was purified using GABA_AR α₁ antibodies in HEK-293 cells co-expressing GABA_ARs (α₁β₂γ₂) and WT/mutant α₁ GlyRs, and the co-precipitating proteins were detected by immunoblotting. Inputs are immunoblots of the same protein in cell lysates before Co-IP. Quantification of WT and mutant GlyR α₁ binding to GABA_AR α₁ subunits (n = 3). The data were normalized to the WT group.

(B and C) Endogenous brainstem GlyRα₁ of WT and GlyRα₁ R271Q (B) or S267Q (C) KI mice was purified using GABA_AR α₁ antibodies, and the co-precipitating proteins were detected by immunoblotting. Inputs are immunoblots of the same protein in tissue lysates before Co-IP. Quantification of WT and R271Q (B) or S267Q (C) mutant GlyRα₁ binding to GABA_ARα₁ (n = 3 mice).

(D) Endogenous brainstem GlyRα₁ of WT and GlyRα₁ M287L KI mice was purified using GABA_AR α₁ antibodies, and the co-precipitating proteins were detected by immunoblotting. Inputs are immunoblots of the same protein in tissue lysates before Co-IP. Quantification of WT and M287L mutant GlyRα₁ binding to GABA_ARα₁ (n = 3 mice). The data were normalized to the WT group.

Data are represented as mean ± SEM. *p < 0.05, ***p < 0.001 based on unpaired t tests; ns, not significant (p > 0.05).