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. 2012 Aug 21;31(20):3976–3990. doi: 10.1038/emboj.2012.235

Figure 1.

Figure 1

Biogenesis of the TGN46 containing transport carriers. (A, B) Permeabilized HeLa cells were incubated under conditions indicated. The reaction mixture was centrifuged at low speed (10 000 g for 10 min) and the supernatant was further centrifuged at high speed (100 000 g for 1 h). The high speed pellet was western blotted with antibodies against TGN46, GalT, and calnexin. The input in the figure refers to permeabilized HeLa cells used as starting material for these experiments. (C, D) The permeabilized HeLa cells were pretreated with H89 or PKI and then incubated with rat liver cytosol (Cyt) and an ATP regenerating system. The high speed pellet was western blotted with anti-TGN46. (D) Quantification of TGN46 in the high speed pellet. The average values of three independent experiments are shown (mean±s.d.). The control (Cont) refers to the permeabilized HeLa cells incubated with DMSO, ATP regenerating system and rat liver cytosol. Figure source data can be found with the Supplementary data.