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. 2005 Jun 21;389(Pt 1):57–62. doi: 10.1042/BJ20050213

Figure 2. Expression of the immunoreactive G-6-PT protein and G-6-P transport in microsomes from transfected cell lines.

Figure 2

The different cell lines were transfected for the expression of the G-6-PT protein as detailed in the Experimental section. (A) Cell lysates of HEK-293 (10 μg of protein) were run on an SDS/PAGE 5–20% gradient gel and blotted on to a nitrocellulose membrane. Blots were reacted with antibodies (Ab) against the indicated epitopes. Lanes 1, mock-transfected cells; lanes 2, G-6-PT–FLAG-transfected cells; lanes 3, rat liver microsomes. The sizes of molecular mass markers (in kDa) are shown. (B) Microsomal proteins (40 μg) from COS7 cells were analysed as in (A). Lanes 1, mock-transfected cells; lanes 2, G-6-PT–FLAG-transfected cells; lane 3, rat liver microsomes. The sizes of molecular mass markers (in kDa) are shown. (C) Microsomal proteins (25 μg) from CHO-K1 cell stable clones were analysed as in (A). Lanes 1, mock-transfected cells; lanes 2, G-6-PT-transfected cells. The sizes of molecular mass markers (in kDa) are shown. (D) Uptake of radiolabelled G-6-P by microsomes from G-6-PT-transfected COS7 cells measured in the presence (Δ) or absence (▲) of the G-6-PT inhibitor S3483. As a control, the uptake of G-6-P in mock-transfected COS7 cells was also measured with (○) or without (●) S3483.