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. Author manuscript; available in PMC: 2012 Feb 14.
Published in final edited form as: Traffic. 2010 Feb 27;11(6):827–842. doi: 10.1111/j.1600-0854.2010.01055.x

Figure 2. Expression of non-regulatable GRASP65 mutants enhances Golgi stacking in interphase cells.

Figure 2

A–E) Confocal images of interphase HeLa cells stably expressing indicated GRASP65 constructs using an (tet-on) inducible retroviral expression system. Cells were treated with 1 μg/mL doxycycline for 72 h, fixed and stained for GM130. F–J) Representative EM images of interphase cells expressing indicated GRASP65 constructs. Bar, 0.5 μm. Note that the number of cisternae in the Golgi stacks was increased in cells expressing the mG mutant (H), the GRASP domain (aa 1–201, I) and PDZ1 (aa 1–112, J) compared with cells expressing GFP (F) and wild-type GRASP65 (G). K) Quantitation of (F–J). Results expressed as the mean ± SEM. The number of stacks quantified for each cell line was indicated. Statistical significance was assessed by comparison to the GFP cell line. ***p < 0.001. Bar charts indicating the actual frequency of each value of cisternal numbers are shown in Figure S1. L) Western blot images of cells described in (A–E). Cells were lysed in SDS buffer followed by western blotting for GFP and other indicated proteins. ’End. p65’, endogenous GRASP65.