Skip to main content
. Author manuscript; available in PMC: 2010 Sep 15.
Published in final edited form as: Biochemistry. 2009 Sep 15;48(36):8528–8539. doi: 10.1021/bi900616w

Figure 2. [3H]-TCA uptake activity and membrane expression of TM3 cysteine mutants.

Figure 2

[A] [3H]-TCA uptake activity normalized to relative cell surface expression as described in iMaterials and Methodsi section. Bars represent mean ± S.D. of three separate experiments with p ≤ ** 0.01 and *0.05 respectively, using ANOVA with Dunnett’s post-hoc analysis. [B] Intact transfected COS-1 cells were treated with sulfo-NHS-SS-biotin as described in iMaterials and Methodsi followed by Western Blot processing. Blots were probed with the anti-hASBT antibody (1:15, 000 dilution) followed by horseradish peroxide-linked anti-rabbit immonuglobin (1:2000 dilution). Each blot was probed for the internal plasma membrane marker, α-integrin (150 kDa) and the absence of calnexin (90 kDa) (data not shown), an endoplasmic reticulum membrane protein representing the negative control in the biotinylated fractions. Marker lanes are shown on the left side of the individual blots. Mature glycosylated hASBT visualizes as the 41kDa band while the lower, 38kDa band (not indicated) represents the unglycosylated species.