FIGURE 8: IGP48 is retained in the ER in short stumpy-like cells.

(A) BSF cells expressing IGP48-HA were incubated at 37°C, 20°C (cold-shock) or with pCPT-cAMP for 12 hours. IGP48-HA was visualised with anti-HA antibody and co-stained with anti-BiP antibody. IGP48-HA remains in the ER in short stumpy-induced cells. DNA was visualised using DAPI. All images are captured at the same magnification, scale bar 2 μm.

(B) Surface biotinylation was performed to determine if IGP48-HA reaches the cell surface in short stumpy-like cells. Cells were cultured in vitro at 37°C or 20°C for 12 hours and the biotinylation assay was carried out as described previously. IGP48-HA was detected by Western blot with anti-HA antibody. Blots were stripped and re-probed for an intracellular marker, RabX1 (localises to the ER) and a surface marker, ISG75, which localises to both the surface and endosomal compartments.

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