gabra1  (Proteintech)

Journal: eLife

Article Title: Phenylhydrazone-based endoplasmic reticulum proteostasis regulator compounds with enhanced biological activity

doi: 10.7554/eLife.107000

Figure Lengend Snippet: ( A ) Representative immunoblot (above) of HEK293T cells stably expressing α1β2γ2(R177G) GABA A receptors treated for 24 hr with the indicated AA263 analog (10 µM). Quantification of band intensities (below) for cells treated with AA263 yne or AA263-5 is shown. ( B ) Immunoblot and quantification of γ2 and BiP in lysates prepared on HEK293T cells stably expressing α1β2γ2(R177G) GABA A receptors treated for 24 hr with AA263 yne (10 µM) and/or Ceapin-A7 (CP7; 10 µM). ( C ) Immunoblot showing the intensity of γ2 over time following cycloheximide (CHX)-chase application (0–4 hr, 100 μg/ml) in HEK293T cells transiently transfected with α1β2γ2(WT) receptors (top) and HEK293T cells stably expressing α1β2γ2(R177G) receptor variant treated with vehicle (middle) or AA263 yne (bottom). ( D ) Representative immunoblot showing total γ2 expression in HEK293T cells stably expressing α1β2γ2(R177G) receptor variant treated with indicated AA263 yne analogs (10 µM, 24 hr). One-way ANOVA followed by post hoc Dunnett’s test was used for statistical analysis. *p ≤ 0.05; **p ≤ 0.01; ****p ≤ 0.0001. See also and . Figure 5—figure supplement 1—source data 1. Excel spreadsheet showing raw data used to generate . Figure 5—figure supplement 1—source data 2. Uncropped gels shown in with the same annotations shown in . Figure 5—figure supplement 1—source data 3. Uncropped gels shown in .

Article Snippet: Recombinant DNA reagent , pCMV6 plasmids containing human GABA A receptor α1 , Origene , Uniprot No. P14867-1 , .

Techniques: Western Blot, Stable Transfection, Expressing, Transfection, Variant Assay

Journal: eLife

Article Title: Phenylhydrazone-based endoplasmic reticulum proteostasis regulator compounds with enhanced biological activity

doi: 10.7554/eLife.107000

Figure Lengend Snippet: ( A ) Representative immunoblot (above) and quantification (below) of surface biotinylated γ2 in HEK293T cells stably expressing α1β2γ2(R177G) GABA A receptors treated for 24 hr with 10 µM AA263 yne or AA263-5. Na + /K + ATPase serves as a loading control. ( B ) Representative immunoblot showing surface γ2 expression in HEK293T cells transiently transfected with α1β2γ2(R177G) receptors and treated with indicated AA263 analogs 10 µM, 24 hr. Na + /K + ATPase serves as a loading control. ( C ) Representative evoked inhibitory postsynaptic current (eIPSC) traces for α1β2γ2(WT) GABA A receptors and α1β2γ2(R177G) GABA A receptors treated for 24 hr with DMSO, AA263 yne (10 µM), or AA263-20 (10 µM). 10 mM GABA (saturating condition) was applied to the recorded cells to evoke currents. Histograms showing changes in eIPSC peak amplitude ( D ) and peak current density ( E ) for the indicated groups. Band intensities were quantified using ImageJ software, normalized to the DMSO control condition. Each data point is reported as mean ± SEM. One-way ANOVA followed by post hoc Dunnett’s test was used for statistical analysis for A and B. Kruskal–Wallis test followed by post hoc Dunn’s test was used for statistical analysis for D and E. *p < 0.05; **p < 0.01; ***p < 0.001. See also and . Figure 5—source data 1. Excel spreadsheet showing raw data used to generate . Figure 5—source data 2. Uncropped gels shown in with the same annotations shown in . Figure 5—source data 3. Uncropped gels shown in .

Article Snippet: Recombinant DNA reagent , pCMV6 plasmids containing human GABA A receptor α1 , Origene , Uniprot No. P14867-1 , .

Techniques: Western Blot, Stable Transfection, Expressing, Control, Transfection, Software