(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.