Fig. 1.

Intracellular localisation of wt zDHHC2 in PC12 cells and comparison with various control intracellular markers. (A) PC12 cells were co-transfected with a mCHERRY-zDHHC2 plasmid and various constructs encoding GFP-tagged proteins. GFP is used as a marker for the cytosol, zDHHC5 for the plasma membrane and Rab11 for recycling endosomes. The left panel shows the localisation of the wt mCHERRY-tagged zDHHC2 and the middle panel the expression of the co-expressed GFP-tagged proteins. Both are represented in FIRE pseudocolour. The right panel is the merge of the left and middle panels, with the wt mCHERRY-zDHHC2 presented in Magenta and GFP-tagged proteins in Green. Scale bar represents 5 μm. (B) The graph shows the normalized mean ratio ± SEM of endosomal fluorescence of the GFP constructs relative to mCHERRY-zDHHC2 (n = 15 zDHHC2 cells, 16 GFP cells, 17 zDHHC5 cells and 20 Rab11 cells). Statistical analysis (ANOVA) shows a significant difference in the localisation of the various markers compared to zDHHC2 (^⁎⁎⁎, p ≤ 0.001). The lines represent the upper and lower ratio of endosomal enrichment that can be calculated (dashed line, Rab 11 vs zDHHC2, value of 1.36, filled line, zDHHC5 vs zDHHC2, value of 0.576). These maximum and minimum levels have been added to every subsequent graph in order to provide easier comparison between different figures. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)