Expression and purification of a functional heteromeric GABAA receptor for structural studies
Fig 8
Identification of α1-EGFP/β2 receptor expression parameters in mammalian cells.
(A) Diagram of pEG BacMam vector used for generating recombinant baculovirus to infect mammalian cells. (B) Addition of 10 mM sodium butyrate to the virus-transduced culture increases receptor yield but induces aggregation. Absolute fluorescence intensities are shown. (C) Infecting cells with a relative MOI ≥ 1 (β:α) increases receptor monodispersity. Area-normalized traces are shown. Cells were grown at 37°C for FSEC traces in (B) and (C). (D) Dropping the temperature of infected cells increases receptor yield and reduces aggregation observed at higher temperatures. Absolute fluorescence intensities are shown in (D).