FIGURE 6. Autophagy is not required for LeuLeu-OMe-induced cell death.

(A, B) Cells were stably transfected with pZJM-TbAtg8.1/8.2 for inheritable and inducible RNAi of TbATG8.1 and TbATG8.2 expression. The cells were induced with tetracycline, and samples were taken at t = 0 h, 24 h, 48 h and 72 h post-induction for immunoblots (A) and cell counting (B). Anti-TbATG8.2 antibody was used to monitor TbAtg8.1/8.2-RNAi efficiency, and anti-PFR2 (a component of paraflagellar rod) was used as a loading control.

(C) Cells induced for TbAtg8.1/8.2-RNAi or not were also treated with 30 µM LeuLeu-OMe, and cell growth monitored as described in Figure 1. Results shown in (B) and (C) are presented as mean ± SD from 3 independent experiments.