Fig. 7. Effect of Rab7 constructs on localisation of endogenous LC3 in HeLa cells. The cells were transiently transfected with pEGFP-Rab7 wild-type (WT), Q67L (constitutively active) or T22N (dominant negative) as indicated on the left, and grown for 1 day. The cells were starved of serum and amino acids for 2 hours, fixed in cold methanol, and LC3 and Lamp1 were stained by immunofluorescence. Optical sections are shown. Overlay of LC3 (green) and Lamp1 (red) is shown in the third column. The fourth column shows GFP fluorescence from the Rab7 constructs. Notice the large ring-shaped LC3-positive structures in Rab7 WT and Q67L expressing cells, indicated by arrows in the LC3 and merge columns. The centre of the rings typically showed labelling for Lamp1 as indicated by the arrows and inserts in the merge column. These structures were not observed in cells expressing GFP-Rab7 T22N. Bar, 20 µm.