Fig. 4.

Starvation and chloroquine treatments increase the extent of colocalization of LC3 with intracellular CX50. (A–P) Confocal images of HeLa-CX50 cells transiently transfected with a plasmid encoding GFP–LC3 and subjected to immunofluorescence using rabbit anti-CX50 and Cy3-conjugated goat anti-rabbit IgG antibodies. Cells were incubated in the absence (A–H) or presence (I–P) of chloroquine (CLQ) for 24 hours in normal growth medium (A–D and I–L) or starved by incubation in HBSS for the final 2 hours (E–H and M–P). The insets in B–D show deconvolved images of cytosolic accumulations co-labeled with GFP–LC3 and anti-CX50 antibodies at higher magnification obtained by spectral two-photon confocal microscopy. Scale bar: 25 μm (A–D), 20 μm (E–P).