Figure 2. Clptm1, Itm2C, and Glg1 Associate with GABA_AR γ2.

(A) For gene ontology analysis, the set of HFY-GABA_ARγ2-associated proteins listed in Table S1 was compared with the entire mouse database to identify pathways and compartments in which these proteins are significantly enriched using the statistical overrepresentation test (PANTHER with Bonferroni correction for multiple testing). Terms with p value <0.001 are listed here.

(B) HEK293 cells were transfected with Clptm1, Itm2C-Myc, or Myc-Glg1 and HA-tagged GABA_AR γ2 alone, GABA_AR γ2 with non-tagged α1 and β2, or HA-CD4 as a negative control. Clptm1, Itm2C, and Glg1 were specifically co-immunoprecipitated with GABA_AR γ2 but not CD4.

(C) HEK293 cells which stably express GABA_ARα1/β2/γ2 were transfected with Clptm1, Itm2C, Glg1, or HA-CD4, each together with GFP to detect the transfected cells for recording. GABA_AR mediated currents were induced by fast application/removal of GABA (1 mM). Clptm1 significantly reduced GABA_AR mediated currents. Clptm1: n=18 cells from 3 independent experiments, Itm2C: n=17 cells from 2 independent experiments, and Glg1: n=21 cells from 2 independent experiments. *** p<0.001, t-test.

(D) Clptm1 was co-immunoprecipitated with GABA_AR γ2 and α1 in whole brain homogenate from transgenic HFY-GABA_ARγ2 mice.

Results are expressed as mean ± SEM.

See also Table S1, Figure S1 and Table S2.