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Figure 2


Fig. 2. In SNX1-suppressed cells, retrograde endosome-to-TGN transport of FITC-STxB is markedly perturbed. HeLa cells were treated with either control or SNX1-specific siRNA. After 72 hours, toxin uptake assays using FITX-STxB (green channel) were performed. Cells were fixed at indicated times and immunolabeled for SNX1 (Alexa594, red channel). In cells treated with SNX1-specific siRNA, the immunofluorescence signal was markedly reduced. (A) When cells were fixed without prior incubation at 37°C, the toxin was found to label the plasma membrane to comparable levels in control and SNX1-suppressed conditions. (B) After 30 minutes of internalization, the toxin showed juxtanuclear enrichment in control siRNA-treated cells, but remained associated with more peripheral structures in SNX1-suppressed cells. Even after 60 minutes (C) and 120 minutes (D), only moderate amounts of the toxin had reached the TGN area in SNX1-suppressed cells with most cells still displaying vesicular enrichment of the toxin (asterisks). Bars, 20 µm.





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