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. Author manuscript; available in PMC: 2010 Oct 1.
Published in final edited form as: Pharm Res. 2009 Aug 14;26(10):2358–2366. doi: 10.1007/s11095-009-9952-9

Fig. 4.

Fig. 4

Percentage Gly-Sar uptake and membrane expression of mutants at G594 of hPEPT1 in transiently transfected HEK293 cells after 72 h post-transfection. A [3H]Gly-Sar uptake (0.5 μCi/ml, 10 min at 37°C) was measured in HEK293 cells transfected with each mutated transporter.*, p<0.05 compared to WT-hPEPT1 uptake. B The membrane localization was visualized with affinity-purified rabbit anti-hPEPT1 primary antibody (1:200) and FTIC-conjugated secondary antibody (1:200). C The HEK293 cells with transfected proteins were biotinylated with sulfo-NHS-LC biotin for 30 min at room temperature. Immunoprecipitation was carried out followed by Western blot analysis using affinity-purified rabbit anti-hPEPT1 primary antibody (1:500) and was visualized using goat-anti-rabbit HRP conjugated secondary antibody (1:10,000) and chemiluminescence.