Figure 3. Inter-Golgi transport of small anterograde cargo.

(A) HeLa cells expressing either ssGFP-FM4-CD8 or ST-RFP (+VSV-G) were mixed and fused. Before fusion, cells were incubated at 20°C for 2 hr in the presence of CHX (100 μg/ml) and AP21998 (500 nM) to trigger the release of the cargo from the ER and its accumulation in the Golgi. Both drugs were maintained during the imaging. For the study of the aggregated cargo (staples), AP21988 was removed 30 min before fusion, and cells were imaged in an AP21988-depleted medium at 20°C. Graphs show quantification of both markers over time for Golgi 1 and 2. Results are representative of three independent experiments. (B) HeLa cells expressing either ssDsred-FM4-hGH or GT-GFP (+VSV-G) were mixed and fused. As for (A) AP21988 was removed to trigger the formation of soluble aggregates within the Golgi. Graphs show quantification of both markers over time for Golgi 1 to 4. Results are representative of two independent experiments.

DOI: http://dx.doi.org/10.7554/eLife.01296.005

Figure 3—figure supplement 1. Inter-Golgi transport is microtubule independent.

HeLa cell expressing GT-GFP (+VSV-G) and ST-RFP were mixed and fused. When required, nocodazole (2.10⁻³ μg/ml) was added 2 hr before the fusion and was maintained during the time course of the experiment. Upper panel, fixed cell fixed 1 hr post-fusion. Lower panel, live imaging of inter-Golgi exchange in nocodazole-treated poly-karyon.

DOI: http://dx.doi.org/10.7554/eLife.01296.006