Fig. 2.

UCH-L1 associates with the endoplasmic reticular membrane. (A) COS-7 cells were transiently transfected with either GFP or GFP-UCH-L1, as indicated. Cells were fixed in paraformaldehyde (PF) or alcohol, and GFP fluorescence was imaged by using identical exposure times and image settings. Alcohol fixation releases cytosolic proteins, including free GFP, from cells. (B) Fluorescence microscopy was used to image COS-7 cells transfected with GFP-UCH-L1 and immunostained with calreticulin antibody (i), as well as SH-SY5Y (ii) and H1299 (iii) cells immunostained with UCH-L1 and calreticulin antibodies. COS-7 and SH-SY5Y cells were fixed in alcohol; H1299 cells were fixed in paraformaldehyde. Green indicates UCH-L1; red, calreticulin; yellow, colocalization; and blue, DAPI nuclear stain. (Scale bar: 10 μm.)