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. Author manuscript; available in PMC: 2020 Dec 1.
Published in final edited form as: Hepatology. 2019 Jun 26;70(6):2156–2170. doi: 10.1002/hep.30772

Figure 2: Subcellular localization of rOATP1A1 and rOATP1A4 when expressed singly and together in the presence or absence of PDZK1.

Figure 2:

A. Human embryonic kidney (HEK) 293 cells were transfected with plasmid encoding sfGFP-rOATP1A1 or mRFP-rOATP1A4. Two days after transfection cells were fixed and stained with primary rOATP1A4 antibody followed by Cy5 secondary antibody and imaged by confocal microscopy with a 60x objective. Expression of sfGFP-rOATP1A1 is in green, mRFP-rOATP1A4 in red, and Hoechst 33342 stained nuclei in blue. Top: Representative images of mRFP-rOATP1A4 transfected HEK293 cells. Bottom: Representative images of sfGFP-rOATP1A1 transfected HEK293 cells. Both proteins are largely intracellular. Scale bars = 10μm. B. HEK 293 cells expressing sfGFP-rOATP1A1 or mRFP-rOATP1A4 were transfected with pFLAG-PDZK1. Cells were fixed and labeled with rOATP1A4 and PDZK1 antibodies. Expression of sfGFP-rOATP1A1 is in green, mRFP-rOATP1A4 in red, PDZK1 in blue, and Hoechst 33342 stained nuclei in gray. Images were taken by confocal microcopy with a 60x objective. Top: Representative images of mRFP-rOATP1A4 and PDZK1 transfected HEK293 cells Bottom: Representative image of sfGFP-rOATP1A1 double transfected with PDZK1 in HEK293 cells. rOATP1A1 but not rOATP1A4 was distributed on the plasma membrane in the presence of PDZK1. Scale bars = 10μm. C. HEK 293 cells were transfected with plasmid encoding sfGFP-rOATP1A1, mRFP-rOATP1A4, and/or pFLAG-PDZK1. Cells were fixed and immunofluorescence labeled with rOATP1A4 and PDZK1 antibodies. Expression of sfGFP-rOATP1A1 is in green, mRFP-rOATP1A4 in red, PDZK1 in blue and Hoechst 33342 stained nuclei in gray. Images were taken by confocal microcopy with a 60x objective. Top: Representative images of mRFP-rOATP1A4 and sfGFP-rOATP1A1 double transfection in HEK293 cells. Bottom: Representative image of triple transfection with mRFP-rOATP1A4, sfGFP-rOATP1A1 and pFLAG-PDZK1 in HEK293 cells. White arrows point to plasma localization of the proteins. Scale bars = 10μm.