Figure 7.

Amino acid starvation-induced autophagy in bloodstream trypanosomes is ATP-dependent. (A) For BSF cells, autophagy was more efficiently induced in cytomix than in gHBSS. The autophagosome associated form II and the un-associated form I of endogenous TbATG8.2 were separated by 12.5% urea-SDS-PAGE and probed with anti-TbATG8.2. The same blots were probed with anti-tubulin as loading control. (B) Cytomix without glucose could not support BSF cell survival. All cells swelled up after 2 h. (C) and (D) YFP-TbATG8.2 BSF cells were cultivated in medium or starved in cytomix supplemented with 1.1, 2.2, 3.3, 4.4 or 5.6 mM glucose for 2 h. The cells were processed for fluorescence microscopy, autophagosome quantification and ATP measurements as described above. Quantification results were shown as mean ± SD of 3 independent experiments.