Fig. 4. GATE16 and GABARAP exist in two forms after processing at their C-termini. (A) Tagged versions of LC3, GATE16 and GABARAP. Positions of Myc and HA epitope tags are indicated. The conserved Gly residues are shown in bold. LC3^{C22, G120A} lacks the C-terminal fragment and Gly120 is replaced with Ala. GATE16^{C1, G116A} and GABARAP^{C1, G116A} were also produced. The C, GA mutants were tagged only with the Myc epitope at their N-termini. (B) Two forms of mammalian Atg8 family are produced. F9 cells transfected with the tagged proteins shown in (A) were cultured under nutrient-rich or starvation conditions for 2 hours and subjected to immunoblot analysis using an antibody against the Myc epitope. Position of processed forms (I and II) and unprocessed forms (pro) are indicated. An uncharacterized form (*) appeared in GABARAP-transfected cells.