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. Author manuscript; available in PMC: 2015 Jul 15.
Published in final edited form as: FASEB J. 2011 Nov 15;26(3):976–986. doi: 10.1096/fj.11-180679

Figure 10.

Figure 10

Immunofluorescence of HeLa SS6 cells transfected with WT hOCT2 tagged with GFP, or with hOCT2-GFP mutants, where every single cysteine of the extracellular loop was substituted by alanine, or with 2 hOCT2-GFP control mutants, where a glycine of the extracellular loop (G120) or a nonloop cysteine (C179) was mutated to alanine. Cellular distribution of the hOCT2-GFP (in green) in the WT hOCT2-GFP, control hOCT2-GFP mutants, and the C63A-hOCT2-GFP mutant is shown. Golgi apparatus (anti-GM30) or ER (anti-calnexin) is labeled in red. Nuclei are labeled with DAPI (blue). Mutation of one single loop cysteine (C63A) led to an incorrect trafficking of the transporter, which is not, or only to a minor extent, expressed in plasma membrane, in contrast with what is observed in WT hOCT2 or the two hOCT2-bearing control mutation-expressing HeLa SS6 cells. Cysteine-mutated hOCT2 accumulates in a cytosolic compartment (plaques) that is not strictly colocalized with ER or Golgi.