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. 2015 Mar 9;16(3):5299–5333. doi: 10.3390/ijms16035299

Figure 4.

Figure 4

Transformation of Golgi stacks after inhibition of the ATP restoration system, coatomer I (COPI), and the SNARE-proteins. (A–D) HeLa cells treated with NZ (5 min) and then BFA + NZ (5 min); (E) GA of HeLa cells, 30 min after microinjection of an anti-βCOP antibody. (A) βCOP (red) and ManII (green) labeling; (B) labeling of β-COP (nano-gold-enhancement, red arrows); (C) Three-dimensional model of the GA; (D) immunoperoxidase labeling of ManII (arrows); (E) Routine EM section; (F–N) Inhibition of membrane fusion or ARF/COPI machine impairs GA shape in permeabilized cells. Ldl F cells were warmed to 40 °C for 6 h (F); SLO permeabilized (G,H); and incubated for 90 min at 32 °C with the ATP restoration system (ARS), GTP, plus native (I,K); COPI-depleted (J,K,N) or ARF-depleted (M) cytosol, with an anti-βCOP antibody (K), partially purified COPI cytosol (K,N), or mARF1 (M), and then prepared for EM. (F,G) Three-dimensional tomographic models; (H,I–M) Routine electron microscopic sections. (N) Immunofluorescence. Arrow in (L) shows the section of COPI vesicle. Bars, 10 µm (A,N); 400 nm (B,H,I–K); 70 nm (C,G); 100 (F); 120 nm (D,E,L,M).