Figure 3.

The dominant-negative effect of GFP-STX17ΔNTD and GFP-STX17FL can be controlled using the tetracyclin-inducible system in HeLa cells. (A) WT and TetON-GFPSTX17ΔNTD and TetON-GFPSTX17FL HeLa cells were cultured with or without DOX (1.5 μg/ml). Two d later, cells were cultured in regular or starvation medium for 1 h and analyzed by immunofluorescence microscopy using anti-LC3 antibody. Scale bars: 20 μm; 2 μm in insets. The experiments were successfully performed twice. (B) WT and TetON-GFPSTX17ΔNTD and TetON-GFPSTX17FL HeLa cells were cultured with or without DOX (1.5 μg/ml) for 3 or 6 d. Subsequently, cells were cultured in regular or starvation medium with or without 100 nM bafilomycin A₁ for 2 h and analyzed by immunoblotting using the indicated antibodies.