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. Author manuscript; available in PMC: 2021 Sep 22.
Published in final edited form as: Pharm Res. 2019 May 15;36(7):101. doi: 10.1007/s11095-019-2634-3

Fig. VI. Increased phosphorylation status of OATP1B1 with V174A variant in the HEK293 stable cell line.

Fig. VI.

A. Phosphorylation of WT- and V174A-OATP1B1. HEK293-FLAG-WT- and –V174A-OATP1B1 cells were seeded at a density of 2-2.5x106 cells/100mm2 dish. Forty-eight hours after seeding, cells were metabolically labelled with 32P-orthophosphate for 5 h at 37°C. After labelling, cells were lysed and whole cell lysates (500 μg) were immunoprecipitated (IP) with FLAG antibody and subjected to autoradiography and subsequent immunoblot (IB) with FLAG and GAPDH antibodies. Representative images from four independent experiments are shown. B. Densitometry of 32P-labelled WT-OATP1B1 and V174A-OATP1B1 was normalized by its respective protein amount, as detected by FLAG immunoblot. A mixed-effect model was used to compare the phosphorylation of OATP1B1 between V174A- and WT-OATP1B1 as described in the Materials and Methods. Model-estimated fold change and associated SE of phosphorylation of V174A-OATP1B1 vs. WT-OATP1B1 is shown (* p<0.05, n=4).