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. 2003 Dec 5;100(26):15560–15565. doi: 10.1073/pnas.2536699100

Fig. 6.

Fig. 6.

Internalization of the HKβ is enhanced by its association with CD63. (A) COS cells transfected with the HKβ alone were biotinylated with biotin-N-hydroxysuccinimide ester and incubated at 37°C to allow internalization. Plates were then cooled to 4°C, and one dish from each time point (in min) was subjected to a MesNa strip. Biotinylated material was collected with streptavidin-conjugated agarose beads. Three independent experiments were performed in triplicate. (B) COS cells cotransfected with the HKβ and CD63-FL were biotinylated with or without the MesNa strip as described above. All cells were lysed in 2% CHAPS and immunoprecipitated with anti-FLAG pAb. The immunoprecipitate was eluted and incubated with streptavidin-conjugated agarose beads. Four independent experiments were performed. The sample from each time point was analyzed by SDS/PAGE and immunoblotting with anti-HKβ mAb. (Right) Signal intensity from each band was quantified by densitometry.