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Figure 6


Fig. 6. The dileucine-like and WE motifs are necessary for intracellular targeting of OA1 in HeLa cells. (A) Immunofluorescence analysis of HeLa cells transiently transfected with expression vectors for mycHis-tagged OA1 wild type (OA1 WT), single mutant (OA1 {Delta}SY and OA1 WE->AA) or double mutant (OA1 {Delta}SY/WE->AA) and immunodecorated with anti-His6 antibodies. OA1 {Delta}SY/WE->AA was colocalized by confocal microscopy with co-expressed farnesylated GFP (EGFP-F), which labels the plasma membrane. In the merge, areas of colocalization can be observed in yellow/orange. Bar, 10 µm. (B) Quantitative immunofluorescence analysis of the subcellular distribution displayed by full-length OA1 constructs in transiently transfected HeLa cells. Following immunodecoration with anti-His6 antibodies, transfected cells were counted and classified as described in Fig. 5A. Results represent the mean ± s.e.m. of 5-10 independent experiments. The total number of cells counted is indicated.





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