Phenotypic characterisation of Δ<i>atg5</i>.

<p>(A) Growth curve of <i>Leishmania</i> promastigotes in HOMEM medium at 26°C. *, Δ<i>atg5</i> differed significantly from WT (p<0.05). (B) The occurrence of GFP-ATG8 puncta in promastigotes after incubation in nutrient-deprived (PBS, ND) and nutrient-rich (HOMEM medium, NR) conditions for 2 h at 26°C. Scale bar, 10 µm. (C) Occurrence of GFP-ATG8 puncta in promastigotes when incubated in the conditions detailed in (B). Means ± SD from four independent experiments. * and **, occurrence of GFP-ATG8 puncta in Δ<i>atg5</i> were significantly different from in WT in nutrient-deprived and nutrient-rich conditions (p<0.05). (D) Western blot analysis of extracts of promastigotes expressing GFP-ATG8 at logarithmic growth under standard conditions and probed with α-GFP antibody. The faster migrating, lipidated band is labelled GFP-ATG8-II while the un-lipidated band migrating more slowly is labelled GFP-ATG8-I.